(A) A total of 16 tissues were isolated from 5 male rats; the tissues
were snap frozen, heat inactivated, homogenized, and solubilized. The extracted
proteins were digested with endoproteinase Lys-C and trypsin, and
lysine-acetylated peptides were enriched by immunoprecipitation. The acetylated
peptide mixtures were fractionated by SCX in a STAGE tip, and three pH elutions
per tissue were analyzed by high-resolution LC-MS/MS on a LTQ-Orbitrap Velos
instrument resulting in identification of a total of 15,474 lysine acetylation
sites from 4,541 proteins.
(B) For liver and muscle samples, results from three technical
replicates prepared from the tissue homogenates are shown. Logarithmized
intensities for acetylated peptides were plotted against each other and shown on
the left side of the diagonal with the corresponding Pearson correlation
coefficients given on the right side of the diagonal. Technical replicates of
the same tissue are highly correlated.
See also Figures S1 and S2.