Verified Rigor RecordRigor Reviewed

Rigor-Reviewed Manuscript

CircHIPK3 promotes colorectal cancer growth and metastasis by sponging miR-7 · 10.1038/s41419-018-0454-8

Name: Verified Rigor Record — CircHIPK3 promotes colorectal cancer growth and metastasis by sponging miR-7 · 10.1038/s41419-018-0454-8
Item: CircHIPK3 promotes colorectal cancer growth and metastasis by sponging miR-7 · 10.1038/s41419-018-0454-8
Rating: 63
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address · Mixed Methods

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: highReviewer agreement: lowEngine: 7.6.0Source: full text

Not verified in this report

No data or code availability links were detected to verify.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

The paper has a well-justified scientific premise and transparent reporting of outcomes and funding, but suffers from significant rigor gaps in study design (no blinding, power analysis, or randomization method), incomplete reporting of biological variables, missing key resource details, and a complete absence of data/code availability. An internal contradiction in the mouse group numbers (32 mice into six groups but only four described) is a critical reporting error.

This evaluation is based on the full published text. Three independent reviewers scored all eight dimensions; their ratings were synthesized with a conservative approach to disagreements. Verification components checked 8 statistical tests (all consistent) and found one retracted reference. The integrity check flagged a high-severity internal contradiction regarding mouse group numbers.

12 major claims checked against the paper's own evidence: all adequately supported.

Recomputed 8 tests: 8 consistent, 0 inconsistent, 8 via agent-written checks.

1 integrity concern flagged (1 high).

Rigor

Eight Dimensions1 pass · 5 warn · 2 fail

12 major claims1 partial11 supported

Prior work on circRNAs and circHIPK3 is cited, including conflicting roles as oncogene or tumor suppressor. The paper identifies the gap in CRC. The hypothesis follows logically from the cited evidence. Limitations of prior studies are addressed by noting the tissue-specific context and the need to explore circHIPK3 in CRC.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The paper clearly establishes its scientific foundation by citing prior work on circRNAs, circHIPK3, and miR-7, and provides a logical rationale for the study.

Evidence

direct quote[Introduction, paragraph 2]

“Zheng et al. showed circHIPK3 was significantly upregulated in liver cancer and promoted cell proliferation, implying that circHIPK3 might be an oncogene. On the contrary, a more recent study found circHIPK3 served as a tumor suppressor to inhibit bladder cancer growth and metastasis.”

direct quote[Introduction, paragraph 3]

“In this study, we identified that circHIPK3 was an oncogene, which was upregulated in CRC and increased circHIPK3 predicted poor prognosis. Furthermore, circHIPK3 could sponge endogenous miR-7 to sequester and inhibit miR-7 activity, thereby leading to increased FAK, IGF1R, EGFR, and YY1 expression.”

direct quote[Introduction, paragraph 2]

“Whether or not circHIPK3 as a suppressor gene or an oncogene still remains under debate.”

PASSReviewer 2· 92% conf

The scientific premise is well-established. Prior work on circRNAs, circHIPK3, and miR-7 in cancer is cited, the rationale linking circHIPK3 to CRC is clear, and limitations of prior work are addressed.

Evidence

direct quote[Introduction, paragraphs 1-4]

“Mounting evidences indicate that circular RNAs (circRNAs) have a vital role in human diseases, especially cancers.”

direct quote[Introduction, paragraph 2]

“However, its role in colorectal cancer (CRC) has not been explored.”

direct quote[Introduction, paragraph 4]

“However, the regulatory roles of circRNAs act as 'miRNA sponges' in CRC are still largely unknown.”

PASSReviewer 3· 95% conf

The paper clearly establishes prior work on circRNAs, circHIPK3, and miR-7, acknowledges discrepancies in the literature, and provides a logical rationale for studying circHIPK3 in CRC.

Evidence

direct quote[Introduction, paragraph 2-3]

“Recently, circHIPK3, a particularly abundant circRNA, have been verified to be involved in metabolic dysregulation and tumorigenesis.”

direct quote[Introduction, paragraph 2]

“Whether or not circHIPK3 as a suppressor gene or an oncogene still remains under debate.”

direct quote[Introduction, last paragraph]

“In this study, we identified that circHIPK3 was an oncogene, which was upregulated in CRC and increased circHIPK3 predicted poor prognosis.”

For animal experiments, random assignment is mentioned but no method is given (e.g., RNG, block). Blinding is not reported for any experiment. No a priori power analysis is provided. Inclusion/exclusion criteria for patients and animals are mostly absent. Outlier handling is not described. The paper does not report independent replication of key findings. Only replicate distinction (three independent experiments) and controls (si-NC, vector, etc.) are adequate.

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitADEQUATE

inclusion exclusionNOT REPORTED

randomization methodINADEQUATE

replicate distinctionADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

The study design lacks key elements of experimental rigor, including randomization method, blinding, power analysis, and explicit inclusion/exclusion and outlier handling criteria.

Evidence

direct quote[Materials and methods, Animal experiments]

“5-week-old male BALB/c nude mice were randomly divided into four groups ( n = 8 for each group).”

absence[Materials and methods, Animal experiments; Results]

Blinding is not mentioned anywhere in the paper.

absence[Materials and methods, Statistical analysis]

No power analysis is reported.

WARNReviewer 2· 85% conf

The study design has moderate rigor. While replication, control use, and some inclusion/exclusion criteria are adequate, key elements like randomization method, blinding, power analysis, and pre-specified outlier handling are not reported.

Evidence

direct quote[Methods, Animal experiments]

“5-week-old male BALB/c nude mice were randomly divided into four groups ( n = 8 for each group).”

paraphrase[Methods, entire section]

“No mention of blinding in any experimental procedure.”

direct quote[Results, several sections]

“Data were represented as means ± S.D. of at least three independent experiments.”

FAILReviewer 3· 85% conf

Major gaps in reporting of randomization, blinding, power analysis, and replication, especially for the animal experiments.

Evidence

direct quote[Methods, Animal experiments]

“5-week-old male BALB/c nude mice were randomly divided into four groups (n = 8 for each group).”

absence[Methods, Animal experiments and Statistical analysis]

Blinding not mentioned

absence[Methods, Statistical analysis]

Power analysis absent

Sex is reported for the animal experiments (male mice) but not for human CRC tissues (demographics table shows gender but with 102 male, 76 female, so sex is reported). Age is reported for human patients (≤65 vs >65). Health status is inferred from the inclusion criteria (no preoperative therapy). Species/strain/source is adequate for cell lines (ATCC) and animals (BALB/c nude mice, source not specified). Housing conditions for animals (light cycle, temperature, enrichment) are not reported. Human demographics are partially reported (age, gender), but race/ethnicity and comorbidities are not reported.

Sub-criteria

demographicsINADEQUATE

sex reportedADEQUATE

sex justifiedN/A

age weight healthINADEQUATE

housing conditionsNOT REPORTED

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 85% conf

Reporting of biological variables is insufficient: sex is not justified for single-sex animal study, age/weight/health are incompletely reported, species/strain sources are missing for animals, housing conditions are not described, and human demographics lack race/ethnicity and comorbidities.

Evidence

direct quote[Materials and methods, Animal experiments]

“5-week-old male BALB/c nude mice were randomly divided into four groups”

direct quote[Table 1]

“Age (years) ≤65 79 41 (51.9%) 38 (48.1%) 0.763 >65 99 48 (48.5%) 51 (51.5%)”

absence[Materials and methods, Animal experiments]

Animal housing conditions are not described.

WARNReviewer 2· 80% conf

Sex is reported for animal experiments but not for human tissue samples; age and health status are partially reported for humans; species/strain/source is adequate; housing conditions for animals are not reported.

Evidence

direct quote[Methods, Animal experiments]

“5-week-old male BALB/c nude mice”

direct quote[Table 1]

“Gender: Male 102, Female 76”

absence[Methods, Animal experiments]

Housing conditions for animals are not described.

WARNReviewer 3· 80% conf

Sex is reported for both human and animal samples, but the use of only male mice is not justified, and important details like weight and housing conditions are missing.

Evidence

direct quote[Table 1]

“Gender: Male 102, Female 76”

direct quote[Methods, Animal experiments]

“5-week-old male BALB/c nude mice”

absence[Methods, Animal experiments]

Housing conditions not reported

The study involves non-public human tissue samples and animal experiments. The paper states approval by the ethics committee of Nanjing First Hospital and written informed consent, which is adequate. Animal experiments were approved by the Animal Care Committee of Nanjing Medical College with acceptance number SYXK20160006. However, regulatory compliance with a named ethical framework (e.g., Declaration of Helsinki) is not stated. This omission makes the dimension a warn, as not all applicable criteria are adequate.

Sub-criteria

iacuc statementADEQUATE

informed consentADEQUATE

irb ethics statementADEQUATE

regulatory complianceNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Ethics approval and informed consent statements for human and animal studies are present and adequate, but regulatory compliance (e.g., Declaration of Helsinki) is not explicitly stated.

Evidence

direct quote[Materials and methods, Cell cultures and patient tissues]

“This study was approved by the ethics committee of Nanjing First Hospital and written informed consent was obtained from each patient.”

direct quote[Materials and methods, Animal experiments]

“The animal experiments were approved by the Animal Care Committee of Nanjing Medical College (acceptance no.: SYXK20160006).”

absence[Materials and methods]

No explicit statement of regulatory compliance (e.g., Declaration of Helsinki) is found.

PASSReviewer 2· 95% conf

Ethical approvals and consent are adequately reported. The study received ethics committee approval for human tissues and written informed consent was obtained. Animal experiments were approved by an IACUC-equivalent committee.

Evidence

direct quote[Methods, Cell cultures and patient tissues]

“This study was approved by the ethics committee of Nanjing First Hospital and written informed consent was obtained from each patient.”

direct quote[Methods, Animal experiments]

“The animal experiments were approved by the Animal Care Committee of Nanjing Medical College (acceptance no.: SYXK20160006).”

WARNReviewer 3· 90% conf

Ethics approval and informed consent are reported for human tissues, and animal ethics is approved with a protocol number, but no explicit compliance statement with a recognized framework (e.g., Declaration of Helsinki) is provided.

Evidence

direct quote[Methods, Cell cultures and patient tissues]

“This study was approved by the ethics committee of Nanjing First Hospital and written informed consent was obtained from each patient.”

direct quote[Methods, Animal experiments]

“The animal experiments were approved by the Animal Care Committee of Nanjing Medical College (acceptance no.: SYXK20160006).”

Antibodies are reported with vendor and catalog numbers (e.g., anti-FAK #3285, Cell Signaling Technology). Cell lines are sourced from ATCC, but authentication (STR profiling) is not mentioned. Mycoplasma testing is stated ('All cell lines were free of mycoplasma contamination') but the method and result are not described. Organisms are identified (BALB/c nude mice) but source is not specified. Key reagents like siRNAs and plasmids are named (Gene-Pharma, Geenseed Biotech) but catalog numbers or RRIDs are not provided. Software tools are identified with versions (SPSS 22.0, GraphPad Prism 6.0, Originpro.9.0, Flowjo V10).

Sub-criteria

mycoplasma testingINADEQUATE

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

Key resources are inadequately identified: antibodies lack dilution information, cell lines are not authenticated, reagents lack catalog numbers, and the animal strain source is not provided.

Evidence

direct quote[Materials and methods, Western blotting]

“The primary antibodies used were anti-FAK (#3285, Cell Signaling Technology), anti-IGF1R (#ab39675, Abcam), anti-EGFR (#4267, Cell Signaling Technology), anti-YY1 (#66281-1-Ig, Proteintech), anti-GAPDH (#ab181602, Abcam).”

paraphrase[Materials and methods, Cell cultures and patient tissues]

“Cell lines were purchased from ATCC, but no STR profiling or authentication is mentioned.”

direct quote[Materials and methods, Oligonucleotide transfection]

“si-circHIPK3, si-c-Myb, miR-7 mimics, and their respective control oligonucleotides were synthesized by Gene-Pharma (Shanghai, China).”

WARNReviewer 2· 85% conf

Key resources are partially identified. Cell lines are sourced (ATCC), antibodies are identified with vendor and catalog numbers, and software tools are named. However, mycoplasma testing is mentioned but method/result insufficient, and reagents (siRNAs, plasmids) lack full identification.

Evidence

direct quote[Methods, Western blotting]

“anti-FAK (#3285, Cell Signaling Technology), anti-IGF1R (#ab39675, Abcam), anti-EGFR (#4267, Cell Signaling Technology), anti-YY1 (#66281-1-Ig, Proteintech), anti-GAPDH (#ab181602, Abcam)”

direct quote[Methods, Cell cultures and patient tissues]

“All cell lines were free of mycoplasma contamination.”

direct quote[Methods, Statistical analysis]

“SPSS 22.0 (IBM, SPSS, Chicago, IL, USA) and figures were produced using GraphPad Prism 6.0 or Originpro.9.0”

WARNReviewer 3· 80% conf

Antibodies are listed with vendors and catalog numbers but lack working dilutions; cell line authentication is not reported; mycoplasma testing is stated but without method details; mouse source is missing; reagent catalog numbers are often absent; software is adequately identified.

Evidence

direct quote[Methods, Western blotting and Immunohistochemistry]

“anti-FAK (#3285, Cell Signaling Technology), anti-IGF1R (#ab39675, Abcam), anti-EGFR (#4267, Cell Signaling Technology), anti-YY1 (#66281-1-Ig, Proteintech), anti-GAPDH (#ab181602, Abcam)”

direct quote[Methods, Cell cultures and patient tissues]

“All cell lines were free of mycoplasma contamination.”

absence[Methods, Cell cultures and patient tissues]

Cell line authentication not reported

Tests are named: Student's t-test, one-way ANOVA, Kaplan-Meier with log-rank, Cox proportional hazards, Pearson correlation. Exact p-values are reported for many comparisons (e.g., p = 0.009 for multivariate HR) but some are reported as thresholds (p < 0.05, p < 0.001). Assumptions (normality, equal variance) are not mentioned or verified. Effect sizes with confidence intervals are reported for the Cox model (HR = 2.75, 95% CI 1.74–6.51) but not for other comparisons. Statistical software is identified. Data presentation: individual data points are shown in figures, but error bars (SD) are defined. A mathematical plausibility issue: In Table 2, the multivariate analysis for Distant metastasis shows HR = 2.24, CI 1.16–4.75, p = 0.044. The CI does not cross 1, so p should be <0.05, which is consistent. However, the CI and p are plausible. There are no demonstrable arithmetic errors based on reported numbers.

Sub-criteria

tests namedADEQUATE

exact p valuesINADEQUATE

effect sizes ciINADEQUATE

data presentationADEQUATE

software identifiedADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 75% conf

Statistical analysis is partially adequate: tests are named and software is identified, but assumptions are not verified, exact p-values are often missing, effect sizes/CI are not reported for most comparisons, and data presentation lacks individual data points.

Evidence

direct quote[Materials and methods, Statistical analysis]

“Differences between the different groups were tested using the Student’s t -test or one-way ANOVA.”

direct quote[Results, Table 2]

“high circHIPK3 expression was an independent prognostic factors for poor survival of CRC patients (hazard ratio [HR] = 2.75, 95% confidence interval [CI] 1.74–6.51, p = 0.009)”

direct quote[Figure legends (e.g., Fig. 1, Fig. 3)]

“Data were represented as means ± S.D. of at least three independent experiments.”

WARNReviewer 2· 80% conf

Statistical methods are partially adequate. Tests are named, exact p-values are sometimes given, and software is identified. However, assumptions are not verified, effect sizes with CIs are limited, data presentation is adequate but could be improved, and there is a mathematical/consistency concern.

Evidence

direct quote[Methods, Statistical analysis]

“Student's t -test or one-way ANOVA. Kaplan–Meier method ... log-rank test. Pearson's correlation coefficients. Cox proportional hazards models.”

direct quote[Results, Abstract]

“hazard ratio [HR] = 2.75, 95% confidence interval [CI] 1.74–6.51, p = 0.009”

absence[Methods, Statistical analysis]

Assumptions of statistical tests are not verified.

FAILReviewer 3· 85% conf

While software is identified and data presentation includes error bars and sample sizes, many key statistical reporting items are missing: test assumptions are not verified, exact p-values are not always given, effect sizes with CIs are only partially reported, and post-hoc tests are not specified.

Evidence

direct quote[Methods, Statistical analysis and figure legends]

“Differences between the different groups were tested using the Student's t-test or one-way ANOVA.”

direct quote[Abstract]

“hazard ratio [HR] = 2.75, 95% confidence interval [CI] 1.74–6.51, p = 0.009”

direct quote[Figure 1 legend]

“* p < 0.05, ** p < 0.01, *** p < 0.001”

The paper does not include any statement about data availability, nor does it deposit raw data (e.g., sequencing, microarray) in a public repository. No accession numbers are provided. Custom code, if any, is not shared. This is a clear failure to meet open science standards.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

No data availability statement, repository deposit, accession numbers, or code sharing are reported, which is a significant omission.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Entire paper]

No repository deposit or accession numbers are mentioned.

absence[Entire paper]

No code sharing is mentioned.

FAILReviewer 2· 90% conf

Data and code availability are not reported. No data availability statement is provided, no repository deposits or accession numbers are given, and code is not shared.

Evidence

absence[Entire paper]

No data availability statement.

absence[Entire paper]

No repository deposits or accession numbers.

absence[Entire paper]

No code sharing.

FAILReviewer 3· 95% conf

No data availability statement, repository deposit, accession numbers, or code sharing are reported. Only supplementary figures are mentioned.

Evidence

absence[Entire manuscript]

No data availability statement

direct quote[After Discussion]

“Electronic supplementary material Supplementary Figures(DOC 5727 kb)”

Methods are described with adequate detail for replication, though some elements (blinding, randomization) are missing. Trial registration is not applicable. No reporting guideline (e.g., ARRIVE for animal studies) is mentioned. All reported outcomes appear to be presented, but there is no pre-registration to verify. Limitations are discussed, including tissue/developmental-stage specificity of circRNA function and the need for further studies. Conclusions are generally proportional, stating circHIPK3 'may have considerable potential as a prognostic biomarker' and 'may be a promising treatment approach,' which is appropriate. Funding and conflicts of interest are reported.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessINADEQUATE

all outcomes reportedINADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

Overall reporting is transparent: methods are detailed, outcomes are reported comprehensively, conclusions are proportional, and funding/COI are disclosed, though a reporting guideline is not referenced and limitations are only briefly discussed.

Evidence

direct quote[Materials and methods]

“Human normal colon epithelial cell (FHC) and colorectal cancer cell lines (HCT116, HT29, SW480, SW620, DLD1) were purchased from American Type Culture Collection (Manassas, VA, USA) and were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Gibco, Vienna, Austria).”

direct quote[Results, paragraph 7]

“The results showed that circHIPK3 status affected the expression of FAK, IGF1R, EGFR, and YY1, but not that of PAX6, XRCC2, and RAF1.”

direct quote[Acknowledgements and Conflict of interest]

“This project was supported by grants from the National Nature Science Foundation of China (Nos. 81472027, 81501820) to S.W. and Y.P.; ... The authors declare that they have no conflict of interest.”

WARNReviewer 2· 85% conf

Reporting transparency is moderate. Methods are described in moderate detail, limitations are discussed, and conclusions are largely proportional. However, no trial registration, no reporting guideline, and no pre-specified outcomes statement.

Evidence

direct quote[Discussion, paragraph 4]

“One likely explanation for these discrepancies is that circRNA have its biological roles in tissue/developmental-stage specific context.”

direct quote[Abstract]

“circHIPK3 may have considerable potential as a prognostic biomarker in CRC, and support the notion that therapeutic targeting of the c-Myb/circHIPK3/miR-7 axis may be a promising treatment approach”

absence[Methods, entire section]

No reporting guideline is mentioned.

WARNReviewer 3· 80% conf

The methods are described but lack some details (e.g., antibody dilutions, exact mouse source); no reporting guideline is referenced; limitations are discussed but not comprehensively; outcomes appear fully reported; conclusions are proportional; funding and COI are stated.

Evidence

direct quote[Acknowledgements and Conflict of interest]

“This project was supported by grants from the National Nature Science Foundation of China (Nos. 81472027, 81501820) to S.W. and Y.P.”

absence[Methods]

No reporting guideline mentioned

direct quote[Discussion]

“Further studies will be required to investigate the role of circHIPK3 in other malignancies.”

17 copyedit issues flagged (1 major): mostly typo, consistency, grammar.

Checked 40 references: 39 verified — 1 retracted.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

As a published paper, this assessment indicates that several methodological and reporting gaps undermine confidence in the reported findings. The critical internal contradiction (mouse group numbers) and the absence of data availability are particularly concerning. An informed reader should weigh these issues carefully; a correction statement addressing the group number discrepancy and addition of data availability information would be warranted.

1.
HIGHother
Correct the internal contradiction in the Results animal experiments section: 32 mice cannot be divided into six groups of 8; the text should state four groups to match the described treatments. Issue a correction to the published article.
This is a demonstrable numerical error that undermines the credibility of the animal experiment reporting and could affect interpretation of the results.
2.
HIGHdata code
Add a data availability statement to the manuscript and deposit raw data (e.g., qRT-PCR Ct values, animal tumor measurements, Western blot images) in a public repository such as Figshare or GEO, with accession numbers.
The complete absence of data availability is a major open science deficit; readers and reviewers cannot verify or reuse the reported findings.
3.
HIGHreporting
Remove or replace the retracted reference (circRNA_100290 in oral cancer, doi: 10.1038/onc.2017.89) with a non-retracted source.
Citing a retracted article is a serious integrity issue; the replacement should provide equivalent evidence if needed.
4.
HIGHrigor
Describe the randomization method used for animal group allocation (e.g., computer-generated random numbers) and specify the randomization unit (mouse).
The current statement 'randomly divided' is insufficient; reviewers and readers need to assess potential selection bias.
5.
HIGHrigor
Report blinding procedures for all outcome assessments (e.g., tumor volume measurement, IHC scoring) or justify why blinding was not performed.
Lack of blinding is a major source of bias in interventional experiments; it is essential for credibility.
6.
HIGHrigor
Provide a sample size justification or power analysis for the animal experiments, specifying the expected effect size, alpha, and power.
The sample size of n=8 per group is not justified; a power analysis would demonstrate that the study was adequately powered to detect meaningful effects.
7.
HIGHrigor
Define pre-specified inclusion/exclusion criteria for patient selection and animal experiments, and describe how outliers were handled in the analysis.
Transparent criteria are necessary to assess potential selection bias and to enable reproducibility.
8.
HIGHstatistics
Report exact p-values and effect sizes with confidence intervals for all cell-based comparisons (e.g., in figures or supplementary tables), not just threshold asterisks.
Threshold-only p-values reduce informativeness; exact values allow readers to assess the precision of the estimates.
9.
HIGHstatistics
Specify the post-hoc test used after one-way ANOVA (e.g., Tukey, Dunnett) in the Methods and figure legends.
Without post-hoc tests, the multiple comparisons within ANOVA are not properly controlled.
10.
HIGHrigor
Provide cell line authentication (e.g., STR profiling) for all human cell lines used, and report the results.
Cell line misidentification is a common source of irreproducibility; authentication is expected for published studies.
11.
HIGHrigor
Report working dilutions and application-specific details for all primary antibodies used in Western blot and IHC.
Antibody dilutions are essential for reproducibility; currently only catalog numbers are provided.
12.
HIGHrigor
Justify the use of only male mice in the animal experiments, or include female mice or discuss the potential sex-specific effects.
Single-sex animal studies should be justified to avoid sex bias in preclinical research.
13.
MEDIUMreporting
Add a statement of compliance with relevant ethical guidelines (e.g., Declaration of Helsinki, ARRIVE guidelines) in the ethics section.
Explicit compliance statements are standardly required by journals and provide assurance of ethical conduct.
14.
MEDIUMreporting
Describe housing conditions for animals (light cycle, temperature, enrichment) in the Methods.
Housing conditions can affect experimental outcomes and are required for reproducibility of animal studies.
15.
MEDIUMreporting
Report the mean and range (or standard deviation) of patient age, as well as race/ethnicity and comorbidities, in Table 1.
Complete demographic characteristics are important for assessing generalizability of the cohort findings.
16.
MEDIUMother
Correct the typo 'Scare bar' to 'Scale bar' in figure legends (Figs. 3, 4, 5, 6).
This is a minor but noticeable error in published figures that should be corrected.
17.
MEDIUMcopyedit
Correct remaining typos: 'reveled' → 'revealed', 'accordiing' → 'according', 'evidences' → 'evidence', 'SigmaeAldrich' → 'Sigma-Aldrich', 'Flowjo' → 'FlowJo', '450 nM' → '450 nm'.
These typos reduce the professional quality of the manuscript and should be fixed in a correction.
18.
MEDIUMreporting
Reference a reporting guideline such as ARRIVE for animal studies in the Methods section.
Using reporting guidelines improves completeness and transparency of the methodology.
19.
LOWreporting
Expand the limitations section to explicitly address the lack of blinding, power analysis, and cell line authentication as potential limitations.
Acknowledging these gaps in the discussion would demonstrate critical self-assessment and guide future studies.

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