Expression levels of BCL2 family members in a panel of BRAF mutant cell lines undergoing BRAF inhibition.

<p>(<b>A</b>) mRNA expression levels of various BCL2 family members were quantified by real-time PCR changes and are plotted as log fold difference from vector control (DMSO). (<b>B</b>) Across our panel of cell lines, <i>BCL2-W</i> and <i>BIM</i> increased significantly from control in the context of BRAF inhibition. PLX4720 (1 µM) was used as BRAF inhibitor.</p

The effect of BRAF inhibition, BH3-mimetics or their combination on cell proliferation, apoptosis and protein expression levels of BCL2 family members in BRAF^V600E melanoma cell lines.

<p>(<b>A</b>) MTT assay demonstrating the effect of BCL-2 inhibition, BRAF inhibition, and their combination, on cell proliferation with their respective combination index (CI) value. (<b>B</b>) Corresponding isobolograms. (<b>C</b>) Fluorescence activated cell-sorting (FACS) for Annexin after indicated drug treatment in a BRAF^V600 cell line, A375. Drug combinations used at a 1∶1 ratio. (<b>D</b>) Western blotting of BIM and MCL1 in a BRAF^V600 cell line, A375 after 2, 6 and 24 hours treatment with a BRAFi, ABT and the combination of both BRAFi and ABT.</p

Protein expression levels of BCL2 family members in patients undergoing treatment with a BRAF inhibitor.

<p>RPPA analysis of tumors from patients with metastatic melanoma shows a significant increase of BID and BIM on BRAFi. (<b>A</b>) Protein expression levels of each gene from pre and on treatment biopsies for each patient are shown as log fold change on treatment. (<b>B</b>) Changes in protein expression levels across patients 10–14 days after initiation of BRAFi are plotted on a log scale as fold change from pre-treatment levels using box and whisker plots (* = P≤0.05).</p

The effect of BH-3 mimetic treatment, BRAF inhibition, and their combination on tumor growth in BRAF V600E mutant xenografts.

<p>(<b>A</b>) A375 xenograft. (<b>B</b>) A2058 xenograft. PLX4720 was used as BRAF inhibitor and ABT-263 was used as BH3-mimetic. Both inhibitors were given to mice PO daily at 100 mg/kg <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101286#pone.0101286-Lee1" target="_blank">[45]</a> for 12 days according to treatment group. Mice were euthanized when tumors reached maximal allowed tumor volume. This occurred between days 9 and 12 for some but not all animals. Error bars represent standard error of the mean (SEM).</p