Fig. 3.

Role of NOX4 on TGF-β-induced apoptosis. Analysis made in Hep3B (Control and CRISPR NOX4) cells. A-B) Representative flow cytometry plots using Annexin V-FITC/PI staining for analysis of apoptosis in Hep3B (Control and CRISPR NOX4) treated with TGF-β for 48h (A) or 72h (B). In A, co-treatment with Q-VD-OPH, supporting the role of caspases in the process. C) Proapoptotic (BMF and BCL2L11) and antiapoptotic (BCL2L1 and MCL1) mRNA expression analyzed by RT-qPCR, normalized to housekeeping gene L32, after TGF-β treatment at 48 h. D) MCL1 and Bcl-xL protein levels analyzed by western blot after TGF-β treatment at 24, 48 and 72 h. β-Actin was used as loading control. In A and C data are Mean ± SD (n = 3–6). *p<0.05 **p<0.01, ***p<0.001. In B and D, a representative experiment is shown.