Fig. 4

Rapamycin inhibited the EMT of HLE-B3 cells cultured under high-glucose conditions by modulating the Notch signaling pathway. HLE-B3 cells were stimulated with 30 mM glucose (HG group), and then treated with DMSO (DMSO group, equal volume solvent of rapamycin), 200 nM rapamycin (RAPA group) for 24 h, respectively. A The expression of E-cadherin, ZO-1, α-SMA and Fibronectin in HLE-B3 cells from each group was detected using Western blotting at 24 h. Statistical analysis of the relative expression of E-cadherin (B), ZO-1 (C), α-SMA D and Fibronectin (E), n = 3. Cell migration after exposure to different treatments for 24 h was detected by performing a transwell assay and scratch wound assay. Representative images and the quantification of the transwell assay (F, G, n = 3, bar = 100 μm) and scratch wound assay (H, I, n = 3, bar = 200 μm). J Expression of proteins in the Notch signaling pathway in HLE-B3 cells at 24 h. Quantification of the relative levels of Jagged1 (K), Notch1 (L), NICD (M) and Snail (N), n = 3. ns indicates P > 0.05; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 and ^****P < 0.0001 vs. NC group; ^##P < 0.01 and ^###P < 0.001 vs. HG group; ^★P < 0.05, ^★★★P < 0.001 and ^★★★★P < 0.0001 vs. DMSO group