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Fig. 6 | Journal of Translational Medicine

Fig. 6

From: Suppression of p66Shc prevents hyperandrogenism-induced ovarian oxidative stress and fibrosis

Fig. 6

p66Shc silencing suppresses oxidative stress and fibrotic factor activation in granulosa cells. Granulosa cells were transfected with p66Shc siRNA followed by treatment with DHT, TGF-β1 or resveratrol. a Levels of p-p66Shc (Alexa Fluor 488) and TGF-β (Cy3) were measured with immunofluorescence (60×). Images are representative of three independent experiments with similar results. Quantification of the p-p66Shc (b) and TGF-β (c) fluorescence is shown. d ROS was assessed by the DCF-DA probe using confocal microscopy (40×). Images are representative of three independent experiments with similar results. e SOD enzymatic activity in granulosa cells was measured by colourimetric method. f The expression of fibrotic factors and AR was assessed by western blot assay. mRNA expression of p66Shc (g), Sirt1 (h), and TGF-β (i) was analyzed by real-time PCR. Three independent experiments were performed with similar results. Data are shown as the mean ± SD. *p ≤ 0.05. DHT dihydrotestosterone, TGF-β transforming growth factor-beta, p-p66Shc phosphorylated 66-kDa Src homology 2 domain-containing protein, ROS reactive oxygen species, SOD superoxide dismutase, Sirt1 sirtuin 1, AR androgen receptor

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