Fig. 4: Nilotinib blocks reactivation of ERK signaling in BRAFi-resistant cells. | Nature Communications

Fig. 4: Nilotinib blocks reactivation of ERK signaling in BRAFi-resistant cells.

From: Combating acquired resistance to MAPK inhibitors in melanoma by targeting Abl1/2-mediated reactivation of MEK/ERK/MYC signaling

Fig. 4

BRAFi (-BR; R) or parental (P) cells were treated with vehicle, nilotinib (nilo; 2.5 μM), PLX4720 (M14 and 451-Lu 1 μM; Mel1617 2.5 μM) or the combination for 24 h, and the resulting lysates probed with the indicated antibodies. Quantitation of key signaling molecules for n = 3–5 independent experiments is shown below blots; green arrows indicate lanes quantified. Mean ± SEM is shown. M14-BR: pMEK n = 5, pERK n = 4, pFRA1 n = 5, MYC n = 4. Mel1617-BR: pMEK n = 5, pERK n = 3, pFRA1 n = 3, MYC n  = 3. 451-Lu-BR: pMEK n = 3, pERK n = 6, pFRA1 n = 4, MYC n = 5. For M14 and Mel1617 cell lines, data are a comparison between PLX + nilotinib vs. PLX lanes, whereas for 451-Lu cell lines, nilotinib alone is compared to vehicle (DMSO). *p < 0.05, **p ≤ 0.01, ***p < 0.001 using one-sample t-tests (two-sided). Actual p values (left→right): a 0.0069, 0.002, <0.0001, and 0.0072. b 0.026, 0.0097, 0.029, and 0.011. c 0.046, <0.0001, 0.00011, and 0.0001. pCRKL (substrate of ABL1/2) is an indirect read-out of ABL1/2 activity and indicates the efficiency of nilotinib-mediated inhibition. For all experiments, BRAFi-resistant lines were maintained in PLX (1 μM), but were cultured for 2 days without PLX prior to plating for experiments.

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