Fig. 3.

Effects of hit compounds on RNA levels and histone marks. a Sybr green real-time PCR mRNA level measurement of EZH2 target genes and executing enzymes following a 48-h compound treatment at different concentrations of MDA-MB-231 cells. Measurements marked with an ‘*’ are below detection limit, most likely due to cell death. All RT-PCR experiments were performed in triplicate, normalised to GAPDH and displayed as fold difference to the untreated sample. Error bars represent the mean ± SD of experiment performed in technical triplicate. b Sybr green real-time PCR measurement of the FBXO32 transcription start site and KRT17 promoter region following chromatin immunoprecipitation, using antibodies to the histone marks shown, of MDA-MB-231 cells treated with three selected compounds at 5 μM for 72 h. Shown are representative examples of triplicate ChIP experiments which consistently showed the same changes. The fold difference to the untreated sample is shown. Each IP-value has been determined as the relative increase to the no-antibody control and then normalised to GAPDH levels