Fig. 3.

NLRC5 deficiency aggravates VSMC proliferation, migration, and dedifferentiation. a Human aortic smooth muscle cells (HASMCs) are transfected with scramble small interfering RNA (siCtr) or NLRC5 small interfering RNA (siNLRC5) for 48 h. HASMC proliferation is measured by MTS assay in the presence or absence of PDGF-BB (10 ng/ml) at the indicated time points. *P < 0.05 vs siNLRC5+PDGF at 36 h after PDGF-BB stimulation. b Edu incorporation (green) is evaluated by fluorescence microscopy. Hoechst 33342 is used as a nuclear stain of HASMCs. Scale bar: 100 μm. c, d After transfection with siCtr or siNLRC5 for 48 h, cell migration is assessed by scratch assay in HASMCs with or without PDGF-BB (10 ng/ml) stimulation for 12 h c. Scale bar: 100 μm. Wound area is analyzed by ImagePro Plus software d. e, f Representative western blotting of NLRC5, PCNA, Cyclin D1, α-SMA, Calponin, Myosin, and Vinculin in HASMCs transfected with siCtr or siNLRC5 in the presence of PDGF-BB for 0, 6 and 12 h (10 ng/ml). Two-tailed Student’s t-test is used to compare two groups d, and analysis of variance (ANOVA) followed by Bonferroni post hoc analysis is used to compare three or more groups a, f. Data are presented as mean ± SD from three independent experiments. *P < 0.05. Original magnification, ×100 (b and c). Source data are provided as a Source Data file