Fig. 2.

Determination of optimal lysate concentrations and test of reproducibility of the HuProt array-based lysate profiling of multiplexed PTM signatures. A, the number of positive hits at increased amount of total tumor lysates. Consideration of the reaction of all four type of PTMs, 50 μg/120 μl lysate each reaction appeared relative high hit numbers. B, Representative cropped images of HuProt array for each PTM assay. For example, Src kinase-associated phosphoprotein 1(SKAP) was found to be readily Tyr-phosphorylated and Protein AMBP was SUMOylated, whereas Annexin A10 (ANXA10) and Glucose-6-phosphate 1-dehydrogenase (G6PD) were strongly ubiquitylated, Peptidyl-prolyl cis-trans isomerase A (PPIA) and Minichromosome maintenance domain-containing protein 2 (MCMD2) were acetylated, respectively, by the lysates tested. C, Four types of PTM reactions on different HuProt arrays showed high reproducibility with TCGA sample #2H4).