Fig. 3: Androgens protect C2C12 cells from senescence induced by H₂O₂.

a–e C2C12 cells were untreated or pre-treated with 10 nM R1881. After 4 h, cells were treated with 150 µM H₂O₂ for 36 h. a The WB of lysates proteins from C2C12 cells, using the antibodies against the indicated proteins are shown. Images are representative of three different experiments. Expression levels of proteins were analyzed by densitometry analysis, using NIH Image J Software. The ratio between cyclin D1/Tubulin (b, left panel) and p16/Tubulin (b, right panel) was evaluated. c Cells were processed for β-galactosidase detection through cytochemistry at pH 6, as described in “Materials and methods” and pictures were acquired. The arrows indicate the blue, senescent cells; bar, 5 µm. d β-gal absorbance was detected by evaluating the absorption at 420 nm of o-nitrophenol. e WB of lysates from C2C12 cells using the antibodies against the indicated proteins are shown. The WB in the upper panel are representative of three different experiments. Expression levels of AR were analyzed by densitometry analysis, using NIH Image J Software. The ratio between AR/Tubulin was evaluated. b, d, e Means and SDs are shown; n represents the number of experiments. The asterisks (*) indicate P < 0.05.