Figure 3

Flow chart of sampling and proteome analysis of human left ventricular biopsies. Human left ventricular (LV) biopsies from patients without (sham) or with remote ischemic preconditioning (RIPC) were lysed in Tris/sodium dodecyl sulfate (Tris/SDS) or in radioimmunoprecipitation assay (RIPA) buffer. Proteome analysis was performed after phosphopeptide enrichment, in-solution digestion, and in-gel digestion with Tris/SDS buffer-lysed biopsies and after in-solution digestion with RIPA buffer-lysed biopsies, respectively. The numbers of all detected phosphopeptides/proteins were displayed in line (a), those with ≥2-fold higher phosphorylation/expression in line (b) and with significant (p < 0.05) ≥2-fold higher phosphorylation/expression with RIPC versus with sham in line (c), as well as those exclusively detected with RIPC or with sham in line (d). The sum of line (b), (c) and (d) was displayed in line (e). All detected phosphopeptides/proteins (line (a)) were subjected to false discovery rate (FDR)-based statistical analysis. Independently of lysis and digestion methods, all proteins detected with a ≥2-fold higher phosphorylation/expression with RIPC versus with sham and those exclusively detected in one group, respectively, were considered (line (f)) for an in-silico pathway analysis.