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. Author manuscript; available in PMC: 2011 Apr 1.
Published in final edited form as: Cancer Res. 2010 Mar 23;70(7):2891–2900. doi: 10.1158/0008-5472.CAN-09-3139

Figure 1.

Figure 1

Inhibition of B-RAF up-regulates FOXD3 levels in melanoma cells. A, WM793 and WM115 cells were transfected with non-targeting control (Ctl) or B-RAF (duplex #1) siRNA. Seventy-two hours post-transfection, cells were lysed and analyzed by Western blotting for FOXD3, B-RAF, phospho-ERK1/2 (pERK1/2) and ERK1/2. B, WM793TR cells expressing either control (LacZ2.1) or B-RAF (hairpins #1, #3, and #7) shRNA were induced (+) with tetracycline or not induced (−) for 6 days. Cells were lysed and samples analyzed by Western blotting as in A. The right-hand panels show A375TR cells expressing either Ctl or B-RAF #1 shRNA −/+ doxycycline for 6 days. C, WM793TR cells expressing either control or B-RAF shRNA were induced (+) with doxycycline or not induced (−) for 5 days. FOXD3 and actin mRNA levels were analyzed by quantitative RT-PCR. Graphed is the average and standard deviation from three independent experiments. The asterisk indicates statistical significance comparing B-RAF shRNA + doxycycline to other conditions (p<0.05). D, WM793 cells were transfected with Ctl, B-RAF#1 or cyclin D1 siRNA. After 72 hours, cell lysates were analyzed by Western blotting for FOXD3, B-RAF, cyclin D1 and ERK1/2.