Figure 4
From: BET bromodomain inhibitors synergize with ATR inhibitors in melanoma
ATRI/BETI combination treatment kills B16 melanoma cells in vitro and in vivo. (a) B16F10-luciferase cells were cultured in RPMI-1640 emented with 10% fetal bovine serum and antibiotics. They were treated with vehicle (0.1% DMSO), 10 μM VE821 (VE), 10 μM RVX2135 (RVX) or indicated combinations. The experiments were repeated twice in biological triplicates. Cells were imaged in a light microscope (a) or counted in a hemocytometer (b). (c) B16F10-luciferase cells were cultured in the presence of vehicle (DMSO), 10 μM of RVX2135 (RVX) and/or the ATRI VE821 (VE; 10 μM) for 48 h and were assayed for viability by adding luciferin (Perkin-Elmer) to a final concentration of 100 μg/ml. Value to achieve synergy is shown with a dotted line. (d) B16F10-luciferase cells were cultured in the presence of vehicle (DMSO), 0.5 μM of iBET762 and/or 2 μM of the ATRI AZ20 for 48 h and were assayed for viability by adding luciferin (Perkin-Elmer) to a final concentration of 100 μg/ml. Value to achieve synergy is shown with a dotted line. (e) Eight 6- to 8-week-old C57BL/6 Albino mice were transplanted subcutaneously with 105 B16F10-luciferase cells. Seven days after transplantation, mice were imaged in an IVIS Lumina III XR machine. After imaging, mice were treated with oral and i.p. vehicle, or oral RVX2135 at 75 mg/kg b.i.d. and i.p. injection of AZ20 at 25 mg/kg for 4 days, followed by imaging again. Shown is the fold change in luciferase signal during treatment (n=4 mice per treatment group)
