Fig. 2.

Global DNA demethylation with the DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-Aza) increases Ca²⁺/calmodulin-dependent protein kinase IIγ (CaMKIIγ) expression in vascular smooth muscle synthetic phenotype (VSM_Syn) cells. Cultured VSM_Syn cells were exposed to 5-Aza (10 μM) for 0 to 5 days (D0–D5). A: CaMKIIγ and CaMKIIδ protein levels were examined by Western immunoblot (IB) using CaMKII isoform-specific antibodies and quantified by densitometric scanning (right). B and C: quantitative PCR was performed to determine Camk2g and Tagln mRNA levels. n = 3–5 analyzed by one-way ANOVA with Dunnett’s post hoc test. D: Rat thoracic aortas were freshly harvested and endothelial-injured or uninjured aortas cultured ex vivo with or without the presence of 10 μM 5-Aza for 5 days. CaMKIIγ protein content was tested by Western blot. n = 3 analyzed by 2-way ANOVA with Tukey post hoc test. *P < 0.05, **P < 0.01, and ***P < 0.001.