Fig. 3

Activation of MAPK pathway mediates JQ1 resistance in HCC. a Western blot analysis of 97-L and 97-H cells treated with 1 μM JQ1 for 48 h. Total lysates were subjected to the indicated antibodies. b Cell viability curves are shown for varying doses of JQ1 with or without 200 nM SCH or AZD in 97-H cells. Cell viability was determined using Cell Titer-Glo at 48 h after incubation. c 97-H cells were treated with vehicle, JQ1, SCH 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. d Western blot analysis of 97-H cells treated with varying doses of JQ1 with or without 200 nM SCH for 48 h. Total lysates were subjectd to the indicated antibodies. Band intensities for MYC were quantified by Image J software and graphed at the right side. e Colony formation assays were performed in 6-well plates. 97-H cells were treated with vehicle, 1 μM JQ1, 200 nM SCH or the combination. 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. Data are presented as mean ± s.d. *p < 0.05