Fig. 6

Validation analysis using quantitative RT-PCR, Western blot analysis, and IHC. A Volcano plot of DEGs associated with the development and differentiation of vascular SMCs (P < 0.05) identified from the RNA-Seq libraries of control and CoA vessel tissues. B qRT-PCR analysis of actinin1, calponin and MYH11 in control and CoA vessel tissues. Real-time quantitative PCR was performed with gene-specific primers. The expression of each gene was normalized with the average expression of the endogenous reference gene β-actin. The error bar indicates the standard error of the mean fold change. C Total protein was extracted from control and CoA vascular tissues for Western blot analysis. D Protein bands were scanned, and relative band intensities were normalized