Fig. 4

JIB-04 inhibited HASMC migration and contractile phenotypes. A-B. The migration of HAVSMCs was measured by transwell assays. HASMCs were treated with DMSO or JIB-04 (0.5 μM) for 24 h, and representative images of migrated cells were obtained under a microscope. Scale bar, 100 μm; Scale bar, 50 μm. Quantitative analysis of the transwell assay (n = 3 per group). C-D. The protein expression levels of MMP2 and MMP9 were detected by western blotting (n = 4 per group). E–F. The contractile phenotype was evaluated by α-SMA immunofluorescence staining, in which nuclei were stained with DAPI (blue) and α-SMA was stained in green (n = 3 per group). Quantitative analysis of α-SMA fluorescence intensity was performed by ImageJ (version 1.8.0). G. Quantification of the mRNA levels of α-SMA, MYH10, Tropomyosin 4, COL1A1, Calponin 1, Caldesmon 1, Fibronectin, and Matrix Gla protein (MGP) in HASMCs after JIB-04 treatment (n = 3 per group). H. Quantification of osteopontin (OPN) and epiregulin mRNA levels in HASMCs after JIB-04 treatment (n = 3 per group). I-J. Western blot showed the protein levels of α-SMA, SM22α, MYH10, and COL1A1 in HASMCs after treatment with DMSO or JIB-04 (0.5 μM) for 48 h (n = 4 per group). K-L. Western blot analysis showed the protein levels of phosphorylated AKT (p-AKT), phosphorylated FOXO3A (p-FOXO3A), and phosphorylated P38 (p-P38) in HASMCs after treatment with DMSO or JIB-04 (0.5 μM) for 48 h (n = 4 per group). β-Actin served as a loading control. *P < 0.05, **P < 0.01, N.S. no significant