Figure 7
From: Cytotoxic unsaturated electrophilic compounds commonly target the ubiquitin proteasome system
Analysis of the phenotypic response to hit compounds. (a) MelJuSo or OPM-2 cells were exposed to compounds as indicated for 6 hours and extracts were processed for immunoblotting. Note the induction of eIF2α phosphorylation and the protein products translated from the spliced form of XBP1 by all compounds in MelJuSo cells and by most compounds in OPM-2 cells. TM = tunicamycin. (b) Western blot analysis of active caspase-3 and (c) PARP cleavage following exposure of OPM-2 cells to test compounds for 6 hours. (d) Analysis of annexin V binding to exposed OPM-2 cells. Cells were exposed to the compounds at 5 μM for 18 hours, incubated with fluorescent annexin V and analyzed by flow cytometry (shown is total annexin V signal). (e,f) Cmap analysis of responses to treatment. MCF7 cells were exposed to the different compounds for 6 hours and gene expression analysed using Affymetrix microarrays. Gene signatures were uploaded into the Cmap data base53. Compounds are compared to pertubational classes and ranked for similarity by Kendall’s tau coefficient. Higher median tau score indicates a high similarity between the compound and the pertubational class of compounds. (e) Ranked similarity in gene expression patterns of MCF7 cells treated with hit compounds compared to gene expression patterns of all cell types in the CMap database, analyzed on the level of pertubational classes. (f) Ranked similarity in gene expression patterns of MCF7 cells treated with hit compounds compared to gene expression patterns of MCF7 cells in the CMap database, analyzed on the level of pertubational classes.
