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. 2017 Jul 17;8(47):82256–82267. doi: 10.18632/oncotarget.19286

Figure 4. Effects of transient versus biased IGF-1R down-regulation on melanoma response to MEK1/2 inhibitor.

Figure 4

(A) DFB, BE and Mel28 melanoma cells were pre-treated with IGF-1R siRNA (48 h), 1 μM Nutlin-3 (12 h), 100 ng/ml CP (12 h) or mock (DMSO) treatment, followed by 24 h of 20 μM MEK1/2 inhibitor U0126 alone. Lysates were analyzed by WB for levels of IGF-1R, p53, phosphorylated (p) ERK1/2, and GAPDH as a loading control. (B) Parallel samples, receiving the same sequential treatment as in (A), were analyzed for total cell number by PrestoBlue fluorescence 48 h after MEK1/2 inhibition. Cell number after U0126 (single agent) treatment is shown as blue area. Predicted sensitivity of combined treatments, if additive, is displayed as red line. Observed cell sensitivity to combination treatments is shown as a yellow line. Data displayed as mean ± S.E.M from three independent experiments, expressed as % of mock treated controls. Statistical analysis: Cell number in observed combination treatments compared to predicted. *P<0.05, **P<0.01, ***P<0.001.