Fig. 3: MHY-1685 regulates mTOR and the associated autophagy in hCSCs. | Experimental & Molecular Medicine

Fig. 3: MHY-1685 regulates mTOR and the associated autophagy in hCSCs.

From: Human cardiac stem cells rejuvenated by modulating autophagy with MHY-1685 enhance the therapeutic potential for cardiac repair

Fig. 3

a Protein expression of autophagy markers mTOR, p-mTOR, LC3-I, LC3-II as evaluated by western blotting (*p < 0.05 vs. vehicle). b Autophagy detection with various concentrations of MHY-1685 in senescent hCSCs by CYTO-ID (*p < 0.05 vs. vehicle). c hCSCs were immunostained with anti-LC3 antibody (yellow), anti-LAMP1 (red), and DAPI (blue) to identify the occurrence of autophagy after treatment with MHY-1685. Scale bar: 50 μm. d The proliferative ability of senescent hCSCs (vehicle, 1 µM MHY-1685-treated hCSCs, 1 µM MHY-1685 + 20 µM + CQ-treated hCSCs) measured by a CCK-8 assay after treatment with MHY-1685 + CQ (*p < 0.05 vs. vehicle, #p < 0.05 vs. MHY-1685) e The senescence of senescent hCSCs was verified by counting the SA-β-gal-positive cells after treatment with MHY-1685 + CQ (*p < 0.05 vs. vehicle, #p < 0.05 vs. MHY-1685). f The differential ability of senescent hCSCs into ECs was verified by a tube formation assay after treatment with MHY-1685 + CQ (*p < 0.05 vs. vehicle, #p < 0.05 vs. MHY-1685). g Representative image of the differential ability of senescent hCSCs to differentiate into SMCs after treatment with MHY-1685 + CQ. α-SMA (green), DAPI (blue). Scale bar: 100 µm. h The differential ability of senescent hCSCs into SMCs was evaluated by qRT-PCR (*p < 0.05 vs. vehicle, #p < 0.05 vs. MHY-1685). Data are shown as the mean ± S.E.M.

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