Geng Integrity Report — Silencing NRF2 enhances arsenic trioxide-induced ferroptosis in hepatocellular carcinoma cells (PLOS One, DOI: 10.1371/journal.pone.0322746)

Academic fraud report · Geng Detector

Summary

Verdict: SUSPICIOUS (yellow). A text-only review of the PLOS One article by Huang et al. (2025) on NRF2 silencing and ATO-induced ferroptosis in HCC cells revealed no definitive evidence of data fabrication, but several integrity concerns were identified. Key issues: (1) The secondary antibody is recorded as 'DaianA32723', which is not a recognized vendor prefix; the correct Invitrogen catalog number A-32723 corresponds to Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed, Alexa Fluor 594. (2) The internal control 'Tubulin (Proteintech, 66240)' is recorded ambiguously, with the canonical β-Tubulin antibody being 66031; the listed number is inconsistent with standard reporting. (3) The abstract contains a duplicated conjunction ('and and'), indicating poor copy-editing. (4) The claim that 'almost all NRF2 was located in the nucleus of HepG2 and Huh7 cells under physiological conditions' contradicts the well-established KEAP1-mediated cytoplasmic retention of NRF2 in unstressed cells. Image-reuse, splicing, statistical and output analyses could not be executed without raw figures and numeric data. Confidence is moderate; findings are consistent with sloppy reporting rather than proven misconduct.

Verdict

🟡 SUSPICIOUS — Multiple textual irregularities and one biologically implausible claim, but no image or statistical evidence available to confirm fabrication.

Key findings

Possible reagent record falsification / typo: Secondary antibody listed as "DaianA32723"; no vendor prefix "Daian" is known. The catalog number A-32723 belongs to Thermo Fisher/Invitrogen and denotes Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594. Mis-recording the host species is a non-trivial integrity concern.
Ambiguous internal control: Western blot loading control recorded as "Tubulin (Proteintech, 66240, 1:5000)". The canonical Proteintech β-Tubulin antibody is 66031; the listed number 66240 corresponds to a different target or HRP-conjugate and is inconsistent with standard reporting practice.
Copy-editing failure in abstract: The phrase "…ATO on HCC and and propose a potential therapeutic strategy" contains a duplicated conjunction, suggesting unrefined copy-paste drafting.
Biologically implausible localization claim: The authors state that "almost all NRF2 was located in the nucleus of HepG2 and Huh7 cells under physiological conditions." Under standard KEAP1-dependent regulation, NRF2 is predominantly cytoplasmic and rapidly degraded in unstressed cells; near-complete nuclear localization at baseline is highly atypical and warrants verification against the original immunofluorescence images.

Evidence highlights

Antibody catalog A-32723 (Invitrogen) → Donkey anti-Rabbit, NOT "Daian"; mismatch between reported reagent description and manufacturer record.
Proteintech 66240 vs. canonical β-Tubulin 66031 inconsistency.
DOI 10.1371/journal.pone.0322746; submission Nov 2024, acceptance ~Mar 2025 (~5 months, timeline reasonable).
Duplicate word "and and" verbatim in published abstract.
No raw images, Supplementary numerical tables, or precise p-values were accessible; image-duplication, splicing, last-digit, and statistical abuse tests could not be performed.

Notes

Limitations: Text-only inspection precludes visual and quantitative fraud detection; conclusions on image integrity, statistical anomalies, and data fabrication cannot be drawn.
Recommended follow-up: request original Fig 4 immunofluorescence micrographs and raw Western blot files from the authors; raise the "DaianA32723" reagent discrepancy on PubPeer; consider a formal editorial query to PLOS One before escalating to an institutional investigation.
Current evidence is consistent with careless reporting and inadequate editorial oversight rather than proven systematic misconduct.