Fig. 3

Mst1 inhibition activated autophagy in mBM-MSCs exposed to H₂O₂. The modulated mBM-MSCs were exposed to 200 μM H₂O₂ for 12 h. a Representative TEM photomicrographs showing the formation of autophagosomes containing organelle fragments (arrows). b, c mBM-MSCs transduced with stubRFP-sensGFP-LC3 lentivirus. The yellow puncta indicate autophagosomes, and the free red puncta indicate autolysosomes. At least 8–10 cells per condition were imaged. Quantification represents the ratio of autolysosome/autophagosome in each group. **p < 0.01, ^#p > 0.05. d, e LC3-II/I, p62, Atg14, Beclin1, and Vsp34 expression levels were evaluated by Western blot analysis. β-actin was used as the loading control. Data are expressed as mean ± SD. **p < 0.01, *p < 0.05. H₂O₂, mBM-MSCs exposed to H₂O₂ only; H₂O₂ + Neg, mBM-MSCs/Neg followed by exposure to H₂O₂; H₂O₂ + sh-Mst1, mBM-MSC/sh-Mst1 followed by exposure to H₂O₂; H₂O₂ + sh-Mst1 + 3-MA, mBM-MSCs/sh-Mst1 pretreated with 3-MA followed by exposure to H₂O₂