Fig. 8
UDCA increases ULK1 levels and LC3II/LC3I ratio, and decreases autophagy flux in C2C12 myotubes. C2C12 myoblasts were differentiated for 4–5 days and incubated with 200 μM UDCA. a After 12 h, ULK1 phosphorylation and total levels were detected by western blot analysis, using ULK1 total levels or tubulin as a loading control, respectively. Molecular weight is indicated in kDa. b Densitometric analysis of p-ULK1 (Ser317) and c ULK1 total protein levels were performed. d After 72 h, LC3I and LC3II protein levels were detected by western blot analysis in the absence or presence of chloroquine. β‐actin was used as a loading control. Molecular weight is indicated in kDa. e A densitometric analysis of the LC3II/LC3I ratio was performed. f Autophagic flux was calculated using the subtraction of LC3II levels in the presence and absence of chloroquine [(LC3II + CQ) − (LC3II-CQ)]). The values are shown as a fold of change and expressed as the mean ± SD of three independent experiments (no paired t-test with Welch’s correction, *p < 0.05 with respect to the control group). SD standard deviation, UDCA ursodeoxycholic acid