Fig. 3
Identification of novel targets for the treatment of YST. A Densitometric evaluation of absolute pixel intensities of the 13 most prominent phosphorylation sites in cell lysates from GCT72, 1411H, NOY-1, and MPAF, as measured by the human phospho-kinase array. B Graphical illustration of potential pharmacological inhibitors based on genomic alterations found in at least 50 % of YST-R samples and changes on protein level. C LD50 values (72 h) of GCT72(-R), 1411H, NOY-1(-R), and MPAF upon treatment with the inhibitors selected in (B). Inhibitors showing LD50 values below 5 µM (green) in GCT72 and higher LD50 values in MPAF (5 - 10 µM = yellow, > 10 µM = red) were further evaluated. D Color-coded changes in cell cycle distribution (G1 = light blue, S = yellow, G2 / M = green, mitotic catastrophe = red, changes < 5 % = grey) upon treatment with LD50 (72 h) concentrations for 24 h with indicated drugs, as compared to the solvent control (DMSO) in GCT72(-R), 1411H, NOY-1(-R), and MPAF. E Lollipop graph summarizing relative number of apoptotic cells in GCT cell lines and fibroblast control cells after treatment with LD50 (72 h) concentrations for 48 h with the indicated drugs in comparison to the solvent control. Of note, due to high autofluorescence of Nintedanib, all cell types were treated with LD50 values (72 h) of GCT72. F Densitometric evaluation of relative pixel intensities of the most prominent phosphorylation sites in cell lysates from GCT72 and GCT72-R treated with AZD7762, Danusertib, Nintedanib, OSU-03012, or SNS-314 (24 h, LD50 72 h) in comparison to the solvent control (DMSO), as evaluated by the human phospho-kinase array