Fig. 3

OA represses autophagy in DM1. Representative Confocal images of LysoTracker staining (red, A-H) and LC3 immunodetection (red, I-P) in TDM (A-D, I-L) and immortalized myoblasts (E-H, M-P) treated for 24 h with 1% DMSO or OA at a final concentration of 0.5 or 2 µM and differentiated for 7 days. (Q, R) Quantification of LC3 puncta per square micrometer (n = 20 to 25 images per condition) in TDM (Q) and immortalized myoblasts (R). (S-V) Quantification and representative western blots of immunoreactive bands for LC3 in control and DM1 TDM (S) or immortalized myoblasts (T). (U, V) Representative western blots, with indication of molecular weight sizes to the right in kDa and quantification of total ATG4 and P62 in protein extracts from healthy control (CNT), DM1 TDM untreated or treated with the indicated concentrations and differentiated for 7 days. DM1 cells were treated with 1% DMSO as a control (-) or 0.5 or 2 µM OA for 24 h (n = 3). Each condition was compared to DM1 cells treated with 1% DMSO. The scale bar equals 20 μm. Nuclei were counterstained with DAPI. The bar graphs show mean ± S.E.M. *P < 0.05, **P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001 according to one-way ANOVA test