Fig. 6

PRMT3 knockdown suppresses the methylation of HIF-1α

(A-B) VSMCs were incubated with/without 10 mM β-GP. Whole-cell lysis was collected for co-immunoprecipitation employing anti-PRMT3 antibodies, and the samples were further subjected to western blot analysis using identified antibodies. (C) Quantification of HIF-1α upon IP normalization in VSMCs. (D-E) Protein levels of PRMT3 in VSMCs infected with Ad-shPRMT3 (shown as PRMT3⁽⁻⁾) or Ad-NC shRNA (shown as NC) in the presence of β-GP. (F) Whole-cell lysis was collected for co-immunoprecipitation employing anti-HIF-1α antibodies, and the samples were further subjected to western blot analysis using anti-HIF-1α and anti-Pan-Rme2a antibodies. (G) Protein levels of HIF-1α in aorta of mice. (H) Whole-tissue lysis was collected for co-immunoprecipitation employing anti-HIF-1α antibodies, and the samples were further subjected to western blot analysis using anti-HIF-1α and anti-Pan-Rme2a antibodies. Data is represented as mean ± SD. p < 0.01 vs. β-GP + NC or CKD. β-GP: β-glycerophosphate; HIF-1α: hypoxia inducible factor 1 subunit alpha; NC: negative control; PRMT3: protein arginine methyltransferase 3; VSMCs: vascular smooth muscle cells