Fig. 5.

Sam68 blocked autophagy flux participating in osteogenic differentiation. A, B Autophagic flux detection by LC3-GFP-RFP system of overexpression and silencing of Sam68 with TNF-α intervention. Yellow dots in merged graphic and free red dots were calculated (n = 7). C, D and E LC3 and P62 protein level was tested after Silencing of Sam68 and then treated with autophagy inducer Rapamycin and inhibitor Bafilomycin, relative level was normalized by GAPDH protein expression(n = 3). F, G Runx2, ALP was tested when Rapamycin was added along with TNF-α treatment (n = 3). H, I Alizarin Red S staining and quantification. ns. not significant difference, *p < 0.05, *p < 0.01, ***p < 0.001