Fig. 2

Transcriptome analyses of i-BET858 treatment highlight known BETi-associated pathways as well as unique features. A Principal component analysis (PCA) showing the distribution of data following RNA-sequencing of OVCAR-3 samples treated with i-BET858, i-BET151 and DMSO control for 4 and 24 h. Three biological replicates of each sample were sequenced. B Volcano plots displaying gene expression levels after 4 and 24 h of i-BET858 treatment (1 µM) in comparison with the DMSO control. Grey dots represent transcripts whose expression did not change significantly as a result of treatment, whilst red and blue dots represent transcripts that were up and down-regulated, respectively. C Venn diagram comparisons of differentially expressed genes between i-BET858 and i-BET151 treatments after 4 and 24 h. D Venn diagram comparison of differentially expressed genes between i-BET858 (4 h, 1 µM), i-BET151 (4 h, 1 µM), and JQ1 (0.125 µM, 40 min, GSE77568). E Table summarising expression changes on genes associated with a core BETi response. F Cell lysates of SKOV3, CAOV3, OVCAR-3 and UWB1.289 (UWB) cell lines were subjected to qRT-PCR validation to confirm changes in expression levels of NRG1, p21 and c-MYC targets. G Gene set enrichment pathway analysis of differentially expressed genes after 4 and 24 h of i-BET858 and i-BET151 treatments. H Gene over-representation pathway analysis of differentially expressed genes uniquely affected with i-BET858 treatment. All values represent the mean ± standard deviation (SD) of three biological samples (*P < 0.05, **P < 0.01, ***P < 0.001)