Fig. 1
From: Autophagy regulates the effects of ADSC-derived small extracellular vesicles on acute lung injury
Effects of Atg5-siRNA on autophagy in ADSCs. A, B Representative western blots showing expression of ATG5, LC3, Beclin-1 and p62 in ADSCs. The expression of ATG5 was most effectively inhibited in siATG5 439-transfected ADSCs. C Statistical analysis of LC3, Beclin-1 and p62 expression. siATG5 treatment markedly inhibited expression of LC3-II and Beclin-1 but promoted that of p62 (the relative expression of LC3-II, control, 0.49 ± 0.01 vs. NC, 0.52 ± 0.03 vs. siATG5, 0.32 ± 0.02. the relative expression of Beclin-1, control, 0.53 ± 40.01 vs. NC, 0.55 ± 0.02 vs. siATG5, 0.40 ± 0.03. The relative expression of p62, Control, 0.62 ± 0.01 vs. NC, 0.66 ± 0.02 vs. siATG5, 1.01 ± 0.03). D Relative mRNA level of LC3, Beclin-1 and p62. siATG5 treatment decreased the mRNA level of LC3 and Beclin-1, but increased that of p62. (the relative mRNA level of LC3, control, 1.00 ± 0.00 vs. NC, 1.07 ± 0.09 vs. siATG5, 0.35 ± 0.05. the relative mRNA level of Beclin-1, control, 1.00 ± 0.00 vs. NC, 0.95 ± 0.08 vs. siATG5, 0.26 ± 0.06. the relative mRNA level of p62, control, 1.00 ± 0.00 vs. NC, 1.14 ± 0.08 vs. siATG5, 1.72 ± 0.27). E Transmission electron microscopy images showing characteristic autophagic ultrastructure in the cells. Autophagosomes are indicated by white arrows. F Quantitative analysis of the number of autophagosomes per mm2 of cell cross sections. The number of autophagosomes per mm2 of cell cross section in the siATG5-treated group was significantly lower than that in the control group (the number of autophagosomes per mm2 of cell cross section, control, 0.053 ± 0.008 vs. NC, 0.049 ± 0.007 vs. siATG5, 0.025 ± 0.006). G Fluorescent images of EGFP-LC3 in ADSCs. H Statistical analysis of the number of cells containing LC3-positive puncta in different groups. The number of cells containing LC3-positive puncta in LPS treatment group was much greater than that in control group, and the number of puncta was decreased by siATG5 treatment (the number of cells containing LC3-positive puncta (%), control, 14.77 ± 2.44 vs. NC, 14.40 ± 3.56 vs. siATG5, 5.40 ± 0.95). The results are expressed as the mean ± SD of three independent experiments. ADSC-NC represents ADSCs transfected with negative control siRNA. ADSC-siATG5 represents ADSC transfected with siATG5