Figure 5. Alterations in the global protein pattern in scAAV6-amiR155-PLBr and scAAV6-shPLB transduced cardiomyocytes assessed by shot gun proteomics and validation by Western Blot.
(A) Scatter plot of protein intensities of cardiomyocytes (CM) 14 days after transduction with 25×103 vg/c of scAAV6-shPLBr (shPLBr), scAAV6-amiR155-PLBr (amiR155-PLBr) or the control vectors scAAV6-shCon (shCon) and scAAV6-amiR155-Con (amiR155-Con). Each dot represents one protein. Red dots represent proteins displaying significantly different levels in comparison to control experiment (fold change >1.5, p<0.01). PLB is phospholamban protein. (B and C) Western Blot analysis of STAT1 and STAT3. CM were transduced and analyzed at 14 days after transduction as in A. (B) Quantitative analysis of three bioreplicates per assay. STAT-specific signal intensities were normalized to GAPDH as loading control. (C) Western Blot images of total and phosphorylated STAT1 and STAT3 were analyzed on 3 separate membranes. GAPDH was detected as a house keeping protein on each membrane. *p<0.05 significant intergroup difference as indicated; n.s., not significantly different.