Fig. 3

Rapamycin activated autophagy by inhibiting the mTOR/ULK1 signaling pathway in HLE-B3 cells cultured under high-glucose conditions. Cells were treated with high glucose (HG, 30 mM glucose), high glucose + DMSO (equal volume solvent of rapamycin) and high glucose + 200 nM rapamycin (RAPA) for 24 h, respectively. A The expression of molecules in the autophagy signaling pathway and autophagy marker proteins. Statistical analysis of the relative levels of LC3II/I B and SQSTM1/p62 (C), p-AKT/AKT (D), p-mTOR/mTOR E and p-ULK1/ULK1 (F), n = 3. G Representative TEM images of autophagosomes (arrows) in different groups. H Quantification and statistical analysis of the number of autophagosomes, n = 5. ns indicates P > 0.05; ^*P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 vs. NC group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 and ^####P < 0.0001 vs. HG ^{group; ★P < 0.05, ★★P < 0.01, ★★★P < 0.001 and ★★★★P < 0.0001} vs. DMSO group