Fig. 7

JIB-04 inhibited autophagic flux by regulating STX17 and RAB7 to disturb the fusion of autophagosomes and lysosomes. A-B. Western blotting was used to evaluate the protein levels of SQSTM1 and LC3 in HASMCs treated with JIB-04 (0.5 μM) for 48 h (n = 4 per group). C. Quantification of the mRNA levels of SQSTM1 and LC3B after treatment with JIB-04 (0.5 μM) for 48 h (n = 3 per group). D-E. HASMCs were transfected with lentivirus harboring tandem fluorescent mRFP-GFP-LC3 to monitor autophagy. Representative images of immunofluorescent VSMCs are shown. Autophagosomes are yellow dots, and autophagolysosomes are red dots. Scale bar: 20 μm. Quantitative analysis of red and yellow dots numbers. *P < 0.05 versus the DMSO group; #P < 0.05 versus the Rapamycin group. F-G. Western blot analysis showed the protein expression levels of STX17, RAB7, LAMP1, and LAMP3 (n = 4 per group). β-Actin served as a loading control. *P < 0.05, **P < 0.01, N.S. no significant