Fig. 3
From: PRMT3 methylates HIF-1α to enhance the vascular calcification induced by chronic kidney disease
PRMT3 is upregulated in the β-GP-induced VSMCs and facilitates the calcification
(A) Experimental protocol of VSMCs. (B) mRNA and protein expression of PRMT3 in VSMCs incubated with 10 mM β-GP for 1 d. 3 d, 7 d, and 14 d. (C) mRNA and protein expression of PRMT3 in VSMCs infected with/without adenovirus Ad-shPRMT3 (shown as PRMT3(-)) or Ad-shNC (shown as NC). (D) mRNA and protein expression of PRMT3 in VSMCs infected with adenovirus, followed by the incubation with/without 10 mM β-GP. (E) mRNA expression of RUNX2. (F) ALP and (G) calcium content. (H-I) Alizarin Red S staining was performed to evaluate the calcification of VSMCs. Scale bar is 100 μm. (J) Western blot bands of α-SMA, SM22α, SMMHC, OPN, OCN. Data is represented as mean ± SD. p < 0.05, p < 0.01 vs. β-GP 0 h, control, NC, or β-GP + NC. ALP: alkaline phosphatase; α-SMA: alpha smooth muscle actin; β-GP: β-glycerophosphate; NC: negative control; OPN: osteopontin; OCN: osteocalcin; PRMT3: protein arginine methyltransferase 3; RUNX2: RUNX family transcription factor 2; SM22α: smooth muscle 22 alpha; SMMHC: smooth muscle myosin heavy chain; VSMCs: vascular smooth muscle cells