Fig. 2

Long-term ex vivo culture causes defective autophagy in human PDLSCs. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 represent significant differences between the indicated columns, while NS represents no significant difference. a Electron microscopy images of P2, P7, and P15 cells. Arrowheads indicate autophagic vesicles. Scale bar = 2 μm. b Presence of autophagosomes (yellow) and their maturation into autolysosomes (red) in adenovirus mRFP-GFP-LC3-transfected cells (P2, P7, P15) with or without Baf treatment (top: representative confocal images, scale bar = 50 μm, ×200 magnification; bottom: quantitative analysis of the percentage of yellow puncta out of the total puncta based on confocal examination). c GFP–LC3 fluorescence levels in P2, P7, and P15 cells before and after Baf treatment (top: representative flow cytometry graphs; bottom: quantitative analysis of the MFI of GFP–LC3). d LC3-II levels in P2 and P15 cells before and after Baf treatment (left: protein levels of LC3-II as determined by Western blot assay; right: quantitative analysis of LC3-II expression in cells). e Expression of cell autophagy-related proteins in P2, P7, and P15 cells as determined by Western blot assay; quantitative analysis of Atg7, p62, and Beclin-1 expression