Fig. 1
From: Phosphoproteomics of short-term hedgehog signaling in human medulloblastoma cells
Scheme of experimental design (a) and analytical workflow. a Three biological replicates of human medulloblastoma cells were treated with DMSO, SAG, vismodegib or EGF in starving medium for 5.0 and 15 min. b Analytical workflow: After treatment, sample preparation, and TMT 10-plex labeling, peptides of each time point and treatment were pooled. Aliquots were utilized for pH 8 fractionation and deep proteome profiling or phosphopeptide enrichment by metal oxide affinity chromatography. Measurements of peptides and phosphopeptides were conducted with reversed-phase HPLC coupled to high-resolution mass spectrometry