Research Paper Volume 12, Issue 8 pp 7534—7548

Protective effects of autophagy and NFE2L2 on reactive oxygen species-induced pyroptosis of human nucleus pulposus cells

Figure 6. Autophagy and NFE2L2 both inhibited CASP1 cleavage. (A) The western blot detecting the expression of SQSTM1, MAP1LC3B and p20 in the nucleus pulposus cells with or without pretreatment with 3-MA before treatment with hydrogen peroxide. (B) The comparison of the data measured in the Figure A. (C) The CCK-8 test revealing the viability of the cells pretreated with different concentration of rapamycin before treatment with hydrogen peroxide (200μM, 3h). (D) The western blot detecting the expression of SQSTM1, MAP1LC3B and p20 in the nucleus pulposus cells with or without pretreatment with rapamycin before treatment with hydrogen peroxide. (E) The comparison of the data measured in the Figure D. (F) The CCK-8 test detecting the effect of ML385 of different concentrations on viability of nucleus pulposus cells. (G) The western blot detecting the expression of NFE2L2 and p20 in the NPCs with or without pretreatment with ML385 before treatment with hydrogen peroxide. (H) The comparison of the data measured in the Figure G. (I) The hochest33342/PI double staining showed the PI positive cells were decreased when nucleus pulposus cells were pretreated with rapamycin and increased when those were pretreated with 3-MA or ML385. (magnification: ×10, scale bar = 200μm) The data were represented as mean ± SEM. *P < 0.05, **P < 0.01.