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B fFigure 2 PHB knockdown inhibits steroidogenesis in gonadotrophin-induced rat GCs. GCs were transiently infected with Ad-shPhb (MOI 5, 10, and 20) or Ad-scrambled (MOI 5, 10, and 20) for 2 h and maintained in culture for 24 h in serum-free media and treated with testosterone (30 ng/mL) in presence or absence of FSH (100 ng/mL) for 48 h in serum-free media. .gl-]g g 6 X  x @`j  BA ?1mj  BA ? t  T$3z ?3  tIndrajit Chowdhury et al. J Mol Endocrinol 2016;56:325-336,;l-];  ; X .  T3z ?D Z $ 2016 The authors*l-]   & X  x 6N  6qљYs ? ̙33ZR___PPT102.]0G+D' = @B D' = @BA?%,( < +O%,( < ++d  5   @ (  @  C ?z7   T3z ? 92    PHB knockdown inhibits steroidogenesis in gonadotrophin-induced rat GCs. GCs were transiently infected with Ad-shPhb (MOI 5, 10, and 20) or Ad-scrambled (MOI 5, 10, and 20) for 2 h and maintained in culture for 24 h in serum-free media and treated with testosterone (30 ng/mL) in presence or absence of FSH (100 ng/mL) for 48 h in serum-free media. (A) Bar diagram represents estradiol (E2) and progesterone (P4) secretions into the medium as measured by RIA. (B) Representative Western blot analyses of protein expression levels of PHB, StAR, p450scc, 3-HSD, and aromatase in GCs induced by FSH under PHB knockdown experimental conditions. Equal amounts of protein (25 g) were applied to each lane. Western blot analyses were analyzed for PHB, StAR, p450scc, 3-HSD, and aromatase. Tubulin was used as an internal control for cytosol. (C) The bar graphs represent the relative percentage change in expression of PHB, StAR, p450scc, 3-HSD, and aromatase as a ratio of the protein levels normalized by tubulin. The bar graphs represent the means.e.m. of results from three independent experiments (n=3). All are significantly different (Pd"0.05) except with same alphabets (a, b, c, and d). 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