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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: Oncogene. 2012 Aug 27;32(31):10.1038/onc.2012.367. doi: 10.1038/onc.2012.367

Figure 4. GAB2 knockdown sensitizes NRAS mutant melanoma cells to apoptosis.

Figure 4

(a) and (b). Melanocytes stably expressing NRASG12D and GAB2WT+NRASG12D were plated onto 3D collagen gels for 24 hours in serum-free medium and analyzed for Annexin-V staining and PI uptake. Experiments were performed in triplicates. Error bars indicate ± standard error. *p<0.05.

(c) MM415 and MM485 are NRAS mutant whereas A2058 and G361 are BRAF mutant melanoma cell lines that overexpress GAB2. These melanoma cells were transfected with control or GAB2 siRNA. Forty-eight hours later cell lysates were analyzed for MCL-1, BCL-2, BCL-XL by Western blotting. Tubulin was used as loading control.

(d) and (e). Forty-eight hours following transfection with control and siGAB2, MM415, MM485, A2058, and G361 cells were replated onto 3D collagen for 48 hours in serum-free medium. Harvested cells were analyzed for Annexin-V staining and PI uptake. Bar graphs indicate percentage of Annexin-V positive cells. Representative traces from one of three experiments are shown. *p<0.05, **p<0.01, and ***p<0.001.