Fig. 3: PRMT5-mediated NF-κB p65 methylation is involved in TMAO-stimulated VCAM-1 expression in VSMC.
From: PRMT5 critically mediates TMAO-induced inflammatory response in vascular smooth muscle cells
A VSMC were pretreated with BAY 11-7085 for 4 h and then stimulated with TMAO (600 μM) for 24 h. Representative blots and quantitative analysis of VCAM-1. The protein expression level was normalized to β-actin. B VSMC were treated with TMAO (600 μM) for indicated periods. Co-immunoprecipitation analysis of the interaction between PRMT5 and p65. C VSMC were treated with TMAO (600 μM) for 24 h. Representative blots of arginine methylation of p65. Immunoprecipitated SYM10 was blotted with an antibody against p65. D Representative blots of arginine methylation of p65. Immunoprecipitated SYM10 was blotted with an antibody against p65 in VSMC transfected with PRMT5 siRNA (siPRMT5) or PRMT5 cDNA plasmid (PRMT5 OE) followed by TMAO (600 μM) stimulation. E Flag-tagged p65 wide-type (p65WT) or R30A mutant (p65R30A) plasmid was transfected to VSMC, and then treated with TMAO (600 μM). Representative blots of arginine methylation of p65. Immunoprecipitated SYM10 was blotted with an antibody against p65. F Flag-tagged p65 wide-type (p65WT) or R30Amutant (p65R30A) plasmid was transfected to VSMC, and then treated with TMAO (600 μM). Representative blots and quantitative analysis of VCAM-1. The protein expression level was normalized to β-actin. Data shown are means ± SD, n = 6 per group from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 using a one-way ANOVA followed by Tukey’s post hoc test.
