Fig. 7

NRF1 regulated SPIDR to protect HCC cells from oxidative stress-induced damage. (A-B), qRT-PCR and Western blotting were employed to measure SPIDR mRNA (A) and protein (B) expression in HepG2 and Hep3B cells transfected with pcDNA3.1 or pcSPIDR. **P < 0.01 vs. pcDNA3.1 group. Ns, non-significant vs. blank group. (C-D), qRT-PCR and Western blotting were employed to measure SPIDR mRNA (C) and protein (D) expression in HepG2 and Hep3B cells transfected with siNC or siSPIDR. **P < 0.01 vs. siNC group. Ns, non-significant vs. blank group. HepG2 and Hep3B cells transfected with siNC, siSPIDR, siNRF1 or co-transfected with siNRF1 and pcSPIDR were used to detect ROS, MDA, SOD and γH2AX levels, as well as cell proliferation. (E-F), quantification of ROS in HepG2 and Hep3B cells without (E) or with (F) H₂O₂ treatment. (G-H), quantification of MDA (G) and SOD (H) in HepG2 and Hep3B cells with H₂O₂ treatment. (I-J), the proliferation of H₂O₂-stimulated HepG2 (I) and Hep3B (J) cells was detected by CCK-8. **P < 0.01 vs. siNC group. &&P < 0.01 vs. siNRF1 group. (K), representative immunofluorescence images of γH2AX foci in HepG2 and Hep3B cells stimulated with H₂O₂. Red, γH2AX. Blue, nuclear. Data for A, C and E-J were shown as mean ± SD of three independent experiments.