Fig. 1

Puerarin directly inhibits OCP autophagy. a The protein levels of Atg7, Beclin1 and Atg5 in the OCPs treated with different levels of Puerarin for 6 h without RANKL were measured by Western Blotting. Compared between Group 0 μM and Group 10 μM, Group 10 μM and Group 25 μM, Group 25 μM and Group 50 μM, respectively, *P < 0.05. b The ratio of LC3II/I in the OCPs treated with Puerarin (50 μM) in the presence or absence of E64d + PEPS A for 8 h without RANKL. c mRNA expression of Atg5, BECN1 or Atg7 in the treated OCPs after infection with lentivirus encoding Atg5, BECN1 or Atg7 (LV- Atg5, LV-BECN1 or LV-Atg5) and the corresponding control vector (LV-Cont). d Viruses-transduced OCPs were treated with Puerarin (50 μM) for 6 h without RANKL, and LC3II/I was detected by Western Blotting. e The autophagosomes (yellow arrows) in the viruses-transduced OCPs treated with Puerarin (50 μM) for 24 h without RANKL were observed using TEM. Scale bar, 1 μm. f Statistical diagrams showed the quantitative results of autophagosomes from E (45 cells from three independent assays). Data are presented as mean ± SEM from three independent experiments. Compared between Group LV-Cont and Group LV-Cont+PR, Group LV-Cont+PR and Groups LV-Atg5/BECN1/Atg7 + PR, respectively, *P < 0.05. PR, Puerarin; Cont, control groups; E, E64D; P, PEPS A