Fig. 5

EGFR inhibition sensitizes HCC cells to JQ1. a Cell viability curves are shown for varying doses of JQ1 in 97-H cells transfected with scrambled shRNA or shEGFR. Cell viability was determined at 48 h after treatment using Cell Titer-Glo. b Colony formation assays were performed in 6-well plates. 97-H cells transfected with scrambled shRNA or shEGFR were treated with 1 μM JQ1 for 6 weeks. After 6 weeks of incubation, colonies were stained with crystal violet and the number of colonies per well was determined and graphed at the right side. c Cell viability curves are shown for varying doses of JQ1 in EGFR-WT or EGFR-I645L expressing 97-L cells. Cell viability was determined at 48 h after treatment using Cell Titer-Glo. d Cell viability curves are shown for varying doses of JQ1 with or without a fixed dose of SCH or ERL in 97-H cells. e 97-H cells were treated with vehicle, JQ1, ERL or the combination for 48 h. Apoptosis was assessed and quantified by Annexin V / PI double staining. Quantification of apoptotic cells was determined based on Annexin V positive cells. f Western blot analysis of 97-H cells treated with 1 μM JQ1, 200 nM SCH, 1 μM ERL or the combination of compounds for 48 h. Total lysates were subjected to the indicated antibodies. Band intensities were quantified by Image J software and graphed at the right side. g Colony formation assays were performed in 6-well plates. 97-H cells were treated with vehicle, 1 μM JQ1, 1 μM ERL or the combination of two drugs. After 6 weeks of incubation, colonies were stained with crystal violet and the number of colonies per well was determined and graphed at following position Data are presented as mean ± s.d. *p < 0.05, **p < 0.01