Research Paper Volume 15, Issue 6 pp 2275—2292

Figure 7. VO-OHpic inhibited oxidative stress induced CEP degeneration and chondrocytes ferroptosis via activating Nrf-2. Chondrocytes were transfected with Nrf-2 siRNA, and treated with TBHP (100 μM) and VO-OHpic (1 μM). (A) Western blot was conducted to examine the protein levels of SOX9, COL2, COL10, RUNX2 and MMP3. (B) The band density of Nrf-2, HO-1, parkin and LC3 was quantified and normalized to control. (C) CEP chondrocytes were treated with TBHP (100 μM) and VO-OHpic (1 μM) for 24 h and western blot was conducted to examine the protein levels of GPX4 and SLC7A11. (D) The band density of GPX4 and SLC7A11 was quantified and normalized to control. (E) Immunofluorescence staining was conducted to examine the expression and localization of GPX4 (red). (F, G) Flow cytometric analysis of endplate chondrocytes stained with Annexin V-FITC/PI. Percentage apoptosis rates were expressed as means ± SD. (H) Chondrocytes were transfected with Nrf-2 siRNA, and treated with TBHP (100 μM) and VO-OHpic (1 μM), western blot was conducted to examine the protein levels of GPX4 and SLC7A11. (I) The band density of GPX4 and SLC7A11 was quantified and normalized to control. Data are presented as mean ± SD from three independent experiments. Scale bar = 20μm. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001.