Fig. 6

Estradiol concentration significantly decreased in female Gm2044 [−/−] mice. (A) The serum estradiol concentration of female lncRNA Gm2044 knockout mice significantly decreased. Serums were isolated from Gm2044[+/+], Gm2044 [+/−] mice and Gm2044 [−/−] mice and then were used to analyze estradiol concentration by ELISA method. (B and C) The NR5A1 protein expression significantly decreased in follicular granulosa cells of Gm2044 [−/−] mice. The protein for follicular granulosa cells of Gm2044[+/+], Gm2044 [+/−] mice and Gm2044 [−/−] mice were isolated and then subjected to western blotting (B) and quantitative analysis (C). (D) The Nr5a1 mRNA expression had no change in follicular granulosa cells of Gm2044 [−/−] mice compared with that in Gm2044 [+/+] mice. The RNA for follicular granulosa cells of Gm2044[+/+], Gm2044 [+/−] mice and Gm2044 [−/−] mice were isolated and then subjected to qPCR. (E) The estradiol level for follicular granulosa cells of Gm2044 [−/−] mice significantly decreased. Culture medium for follicular granulosa cells of Gm2044[+/+], Gm2044 [+/−] mice and Gm2044 [−/−] mice were used to analyze estradiol concentration by ELISA method. (F and G) Knockdown of EEF2 can reverse the elevated effects of Gm2044 on NR5A1 protein level in follicular granulosa cells of Gm2044 [−/−] mice. The protein was isolated from Gm2044 [−/−] mouse follicular granulosa cells transfected with indicated plasmid and siRNA, and then subjected to western blotting (F) and quantitative analysis (G). (H) Knockdown of EEF2 can reverse the elevated effects of Gm2044 on estradiol concentration in follicular granulosa cells of Gm2044 [−/−] mice. Culture medium for Gm2044 [−/−] mouse follicular granulosa cells transfected with indicated plasmid and siRNA were used to analyze estradiol concentration by ELISA method. si-EEF2, siRNA for EEF2 () and siRNA for negatice control (si-NC) *, p < 0.05