Figure 6

Validation of the most representatitve protein changes detected in remote myocardium after acute infarction. (a) Western blot analysis and quantitative densitometry of TPM1 confirming the transient increase in thin filament sarcomeric proteins early after reperfusion. Full-length blots are presented in Supplementary Figure S8 as well as their raw integrated densities before and after normalization can be consulted in Supplementary Figure S12. Protein values are normalized relative to GAPDH. Results are shown as the mean ± s.e.m. of values obtained in at least four animals. *Indicates p value < 0.05 versus baseline by Mann-Whitney test. (b) Label free proteomics analysis confirming the decrease in thick filament proteins and an increase in thin filament proteins early after reperfusion (in at least four animals). Results are expressed as in Fig. 4d. Label free individual protein quantifications included in the thick filament and thin filament protein groups are shown in Supplementary Figure S14. (c) Quantitative analysis by PRM targeted proteomics of proteins ANT2, ACO2, ATP5A1, ATP5O, UQCRC1, MDH2, COX6B, GAPDH, MYH7, MYOM1, MYL3 and TPM1, confirming the transient changes in abundance of mitochondrial and contractile sarcomeric proteins after early reperfusion. Protein values are normalized by the total base peak area of the chromatograms. Results are shown as the mean ± s.e.m. of at least four animals at each time point in two technical replicates. *Indicates p value < 0.05; **indicates p value < 0.01, ***indicates p value < 0.001 versus baseline by Mann-Whitney test. iTRAQ quantifications of the proteins from panels a and c are shown in the Supplementary Figure S11b for comparison.