Fig. 2

Reduced granulosa cell mir-379-5p content is associated with increased PDK1-mediated cell proliferation in an androgenized PCOS rat model. Lower cellular mir-379-5p content in dihydrotestosterone (DHT)-induced PCOS rat model was associated with increased granulosa cell protein content of PDK1 and P-PDK1, and P-PDK1/PDK1 ratio. Increased PDK1 protein content was accompanied by activation of Akt pathway (increased P-Akt/Akt ratio) and increased granulosa cell proliferation (higher MCM2 protein content). Granulosa cells were isolated from 21 day old rats randomly divided into two experimental groups (Non-PCOS and PCOS), and subcutaneously implanted for 28 days with a capsule without and with DHT respectively, as per Materials and Methods. Protein extraction and Western blotting were performed as described previously [15]. Signal intensities generated on the film were measured densitometrically using Image J and normalized over that of GAPDH. U6 RNA was used to normalize miR-379-5p content in granulosa cells. Results are expressed as means ± SEM (n = 3 replicates, each from 1 rat). Data were analyzed by Student “t” test. *P < 0.05, **P < 0.01