Modulation of TMZ-mediated DNA damage signaling by onalespib in patient-derived glioma stem cells and the effect of onalespib on transcriptome and proteome in patient-derived glioma cells. (A) Immunoblot analysis of effect on levels of p-ATM, p-ATR, and p-CHK1 (as measures of DNA damage signaling) and cleaved PARP (reflecting induction of apoptosis) in GSC811, GSC11, GSC262, and GSC23 cells after TMZ treatment and suppression of this effect by pretreatment with onalespib (0.4μM). GAPDH was loaded as a control and HSP70 assayed as a measure of HSP90 inhibition. Figures are representative of three independent experiments. (B) RNA-seq data analysis of expression of various classes of genes on GSC811 and GSC262 exposed to onalespib (0.4 μM for 24h) compared with untreated control cells. Upregulated (red) and down regulated (green) genes are indicated. (C) Changes in select set of proteins in GSCs exposed to onalespib assessed by the RPPA assay.