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. 2021 Feb 16;9:626927. doi: 10.3389/fcell.2021.626927

FIGURE 5.

FIGURE 5

FSH degrades lipid droplets by enhancing autophagy to promote progesterone secretion in porcine follicle GCs. (A) Porcine adherent granulosa cells were transfected with Atg5 siRNAs (siAtg5 1-3#), NC siRNA, and blank control (Mock) for 24 h and then harvested and analyzed by immunoblotting with antibodies against Atg5, LC3, and actin. (B) Porcine adherent granulosa cells were transfected with Beclin1 siRNAs (siBeclin1 1-3#), NC siRNA, and blank control (Mock) for 24 h, harvested, and analyzed by immunoblotting with antibodies against Beclin1, LC3, and Histone 3. (C) BODIPY 493/503 LD staining of adherent granulosa cells, pre-treated with OA for 12 h, after treatment with NC siRNA, siAtg5 2#, and siBeclin1 3#, and then treated with FSH for 24 h; Hoechst 33342 staining of the cell nuclei, Bars: 25 μm. (D) The average number of LDs per granulosa cell in the Mock, NC, Mock + FSH, siAtg5, siAtg5 + FSH, siBeclin1, and siBeclin1 + FSH groups. (E) The average size of lipid droplets per granulosa cell in Mock, NC, Mock + FSH, siAtg5, siAtg5 + FSH, siBeclin1, and siBeclin1 + FSH groups. NC, negative control; NS, no significant. (F) The relative progesterone level in the culture medium from mock, NC, mock + FSH, siAtg5, siAtg5 + FSH, siBeclin1, and siBeclin1 + FSH groups was calculated as pg/μg protein. *, P < 0.05, **, P < 0.01.