Fig 2: UHRF1 depletion deregulates redox‐responsive genes in RB cells. (A) Heat map of differentially expressed genes related to redox and oxidative stress in shUHRF1 Y79 cells. Each column represents an independent replicate. (B) qRT‐PCR analysis for the relative expression of indicated genes in Y79 shCTL and shUHRF1 cells. The bar graph is shown as the mean ± SD of fold changes from four independent experiments, relative to the normalized level in shCTL cells. **P < 0.01, ****P < 0.0001: unpaired Student's t‐test (two‐tailed). (C) Basal expression changes in GSTA4 and TXN2 upon UHRF1 knockdown in Y79 cells. (D) Immunoblots for indicated proteins in Y79 shCTL and shUHRF1 cells after exposure to 1 µm MS‐275, 1 µm SAHA, and 1 mm NaBu for 24 h. (E) Immunoblots in Weri‐Rb1 shCTL and shUHRF1 cells treated with 1 µm MS‐275 or 1 mm NaBu for 20 h.

Fig 3: Representative fluorescence micrographs show (a) peroxiredoxin 5 (PRX5); (b) peroxiredoxin 6 (PRX6); (c) sulfonylated peroxiredoxin peptide (PRX-SO3) positivity in the cornu ammonis 1 (CA1), CA3, and gyrus dentatus (GD) of the hippocampus and the primary motor cortex (M1) area of rat brains (5 samples per each group) subjected to the IR and IPC. White arrows indicate positive cells and red arrows are PRX6 positive astrocytes. Nuclei were labeled with blue DAPI. Scale bar = 50 μm; (d) Western blots for PRX5, PRX6, PRX-SO3, and thioredoxin 2 (TRX2) detected in IR and IPC groups. Results in the bar graphs are presented as the relative protein content normalized to β-actin and compared to the basal control value. Graphs show the mean ± SEM for at least 3 replicates. A one-way ANOVA with post hoc comparisons by the Student–Neuman–Keuls test was carried out to test for significant differences when compared to the control. CON (control), IR (ischemia-reperfusion), and IPC (ischemic preconditioning).

Fig 4: A schematic illustration depicting the mechanism by which disulfiram(DSF) exerts its inhibitory effect on BRAFi-resistant cells by inducing mitochondrial dysfunction.DSF disulfiram, TXNIP thioredoxin-interacting protein, TRX2 thioredoxin 2, MT-ROS mitochondrial reactive oxygen species, OCR, oxygen consumption rate, ΔΨm mitochondrial membrane potential, BRAFi BRAF inhibitors.

Fig 5: TXNIP provokes mitochondrial oxidative stress in vemurafenib-resistant cells via the interaction with TRX2.A Interactome of TXNIP by the STRING database. B The volcano plot of proteins interacting with TXNIP by Data-Independent Acquisition (DIA) quantitative proteomics. C The heatmap of the proteins interacting with TXNIP by Data-Independent Acquisition (DIA) quantitative proteomics. D CCK-8 assay of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. E Colony formation of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. F TEM observation of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. G Immunofluorescence to determine MMP of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. H Seahorse assay of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. I ATP levels of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. J Flow cytometry to detect mt-ROS of TXNIP-in-1 pretreated vemurafenib-resistant cells exposed to Vem or Vem+DSF. Data represent the mean ± SD of triplicate. The differences were analyzed using one-way ANOVA. *P < 0.05, **P < 0.01, and ***P < 0.001. ns non-significant.