Fig. 7: Ablation of ERα normalized gender difference in autophagy and visceral adiposity.

a Knockout (KO) of ERα in sWAT-activated ULK1 by de-phosphorylation at Ser757. b KO of ERα in vWAT-activated ULK1 by de-phosphorylation at Ser757, which diminished the gender difference in ERα expression and p-ULK1 observed in the WT mice. c, d The vWAT in WT females had lower autophagy activity (LC-3II and p62 turnover) than that in WT males (c), but KO of ERα abolished the gender difference (d). The representative Western blotting images were presented in Figs. 4s and 5s. In densitometric analysis, the band densities of investigated proteins were normalized against that of GAPDH, and the fold changes were calculated by taking the normalized density of male group as “1”. For autophagy flux, we first normalized the band densities of LC3-II and p62 against that of GAPDH, then calculated the differences of normalized densities in the presence vs. the absence of autophagy inhibitor; lastly, the differences were shown as fold changes by taking the male group as “1”. e–h WT males had higher vWAT mass than the WT females (f), but KO of ERα abolished the gender difference (h). Overall, KO of ERα increased adiposity in both sWAT and vWAT, presumably due to the enhanced autophagy that promotes adipogenesis. WT wild-type, KO knockout of ERα, M male, F female, WT/M wild-type males, WT/F wild-type females, KO/M knockout males, KO/F knockout females; *p < 0.05; **p < 0.01; n.s. not significant; n = 5–8