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. 2017 Nov 16;11(12):1806–1825. doi: 10.1002/1878-0261.12151

Figure 1.

Figure 1

BRAFi resistance results in metabolic reprogramming toward arginine addiction and circumvention of autophagy due to downregulation of AMPK‐α1. Parental and BR cells were incubated with ADI‐PEG20 (100 ng·mL−1) for 72 h (A) or HCQ alone with or without for 0–72 h (D). Their cell lysates were subjected to immunoblotting. (B) Glucose uptake activity in parental and BR cells was analyzed by 2‐NBDG uptake using flow cytometry (FACS). (C) RNA levels of CAT‐1 and CAT‐2 were detected by qRT‐PCR. On the other hand, MEL‐1220 and A2058 cells were transfected with individual siRNAs (50 nm) against AMPK‐α1, nontargeting (NT) siRNA, or transfection reagent alone (vehicle, Veh). These transfectants were analyzed using immunoblotting (E,F), glucose uptake (G), MTT assay (# P < 0.05 and ## P < 0.01) (H) and qRT‐PCR (I). Data are represented as mean ± SEM (n = 3, *P < 0.05, **P < 0.01, and ***P < 0.005). Fig. 1A reproduced from our previous study (Li et al., 2016).