Fig. 2

Plumbagin reduced the inflammasome overactivation and pyroptotic death of Granulosa cells in PCOS mice. A, B GCs from PCOS mice were isolated and cultured for 48 hours. Inhibitors for different cell death pathways were added as labeled in the figure. Cell survival was quantified with two approaches. n = 4. A. LDH release to the culture medium. B Relative cell viability by alamarBlue™ Cell Viability Reagent staining. C-F GCs were isolated from the ovary of control, PCOS and PCOS mice treated with Plumbagin, the activation of inflammasome was measured. C Activation of Caspase-1 indicated by staining with FLICA probe (Representative graph). D Activation of Caspase-1 indicated by staining with FLICA probe (statistic, n = 4). E. Immunoblot for the amount of active fragment Caspase-1-p10 and GSDMD-N. F. Protein level of Caspase-1-p10 and GSDMD-N normalized with b-actin. n = 3. G. IL-1β level in the culture supernatant were detected by ELISA after 24 hours. n = 5. H. IL-18 release to the culture medium after 24 hours were quantified with ELISA. n = 5. Individual data points are displayed. Data are mean ± SEM. Unpaired one-way ANOVA were used to analysis the difference. *P < 0.05; **P < 0.01; ***P < 0.001