Fig. 3

Rapamycin induced autophagy in Ad-hMSCs. a Transmission electron microscopy verified that Ad-hMSCs treated with rapamycin exhibited numerous cytoplasmic vacuoles, most of which were identified clearly as autophagosomes (with a visible double membrane; red arrow). b mRNA expression of autophagy markers in Ad-hMSCs treated with or without rapamycin was detected by real-time PCR. c Protein extracts from Ad-hMSCs treated with or without rapamycin were analyzed by Western blotting using antibodies targeting phosphor-mTOR, mTOR, phosphor-Rictor, Rictor (a novel binding partner of mTOR), phosphor-Raptor, Raptor, Beclin 1, ATG5, ATG7, and LC3II (left panel). Data in the graphs show the relative intensities of the target proteins normalized to β-actin; values are presented as means ± SD (n = 3). ^* P < 0.05; ^** P < 0.01. Ad-hMSCs adipose tissue-derived human mesenchymal stem cells