Activity

For digoxigenin, digitoxigenin, digoxin, and lanatoside C treatment options. Green box: Evofosfamide Apoptosis Phosphoprofiles of GW8510 treatment options cluster across cell lines, although retaining some cell line-specific responses. B, A summary on the pairwise correlations involving non-self (a sample of a single cell sort treated with 1 compound compared with all other cell line and remedy combinations), the identical drug remedy across cell lines (all biological replicates from the identical drug compared in two different cell lines), as well as the same drug treatment inside each and every cell line are shown (all biological replicates in the exact same drug compared within a single cell line).The robust rightward shift toward 1 demonstrates excellent “replicate recall,” which means that biological replicates are probably to be strongly correlated. Taken together, the middle and bottom panels suggest that there is a robust lineage-specific element to signaling responses in cells although some commonality persists at a lineage-independent level. To further explore the overall performance qualities of the assay, we performed a dose response experiment with staurosporine. A heatmap illustrating the molecular signaturesfrom a subset of P100 probes is shown in Fig. 5A. Though staurosporine is frequently annotated as a broad-spectrum kinase inhibitor, we observed examples of phosphopeptide probes whose levels decrease and others that unexpectedly raise in response with escalating staurosporine dose (Fig. 5A?C). The truth that we observe a diversity inside the responses of probes, exactly where some comply with the anticipated behavior whereas other people demonstrate exclusive behaviors, suggests that we have selected analytes that have informative, dynamic responsesMolecular Cellular Proteomics 15.Reduced-representation Phosphosignature Profilinglog2 fold-change vs. medianA. A.Staurosporine (uM)0.five 10–B.1.four 1.three 1.two 1.1 1.Cluster ProfilesPC3 Cluster1 Profile: Expected Behaviorlog2 fold-change vs. medianCell LinesMCF7 PC0.9 0.8 0.7 0.6 0.five 0.four 0.three 0.2 0.1 0.0 -0.1 -0.two -0.three -0.four -0.5 -0.six -0.7 -0.eight one hundred.5 0.Staursporine(uM) CellType1 0.five 0.5 1 5 five 10 ten 20Gene0.5 0.five 1 1 5 five ten 20 20SiteZNF672 BAT2 WDR20 ATAD2 PFKP KIAA1704 DDX54 RBM17 FASN FASN ZC3H14 HN1 TMPO CASC3 LIMA1 ANLN BAT2D1 GPATCH8 JUND PAK2 KIAA0701 TMPO RPL12 MAP4 PLEC1 ZC3HC1 EIF4A3 ALS2 NUP214 KIF4A C22orf9 SH3KBP1 RPS6KA1 RPS6KA1 RPS6 EIF4A3 MARK2 NOLC1 RBMS189 S1219 S465 S327 S386 S105 S75 S222 S207 S207 S515 S87 T160 S265 S491 S295 T2675 S1035 S100 S2 S935 S306 S38 S2218 T3893 S321 S12 S483 S1012 S801 S304 S230 S230 S230 S235 S12 T596 T611 SStaurosporine [uM]C.1.0 0.9 0.eight 0.7 0.6 0.five 0.4 0.three 0.2 0.1 0.0 -0.1 -0.2 -0.3 -0.4 -0.five -0.six -0.7 -0.8 -0.9 -1.0 -1.1 -1.2 -1.3 -1.four -1.five -1.6 -1.7 -1.8 -1.9 -2.0 -2.1 -2.two -2.3 -2.4 -2.5 1.PC3 Cluster2 Profile: Unexpected Behaviorlog2 fold-change vs. median0.0.Staurosporine [uM]FIG. 5. Signature strength inside the P100 assay is dose responsive. A, A heatmap illustrating a condensed signature derived from a subset of P100 probes as a result of treati.