Fig. 2

IDH1 knockdown suppresses melanoma cell growth and induces ROS under glucose withdrawal. A Sanger sequencing of PCR amplicons correlated with codon 132 of the IDH1 gene in A375 and SK-MEL-28 cells. B qPCR and immunoblot analysis for IDH1 expression in A375 and SK-MEL-28 under 2.5 mM glucose compared with 25 mM glucose for 48 hours. C qPCR and immunoblot analysis for IDH1 expression after IDH1 silencing by siRNA oligos (si.IDH1) compared with control (si.CTRL) in A375 and SK-MEL-28 cells. D Relative ROS levels in si.CTRL and si.IDH1 A375 and SK-MEL-28 cells for 48 hours under the indicated glucose concentrations. E Schematic of the IDH1 enzymatic reaction. F Relative NADPH levels in A375 and SK-MEL-28 cells cultured under the indicated conditions for 72 hours. G Relative GSH levels in si.IDH1 and si.CTRL A375 and SK-MEL-28 cells under the indicated glucose concentration. H Relative 8-OHdG levels in DNA extracted from A375 and SK-MEL-28 cells under indicated conditions for 48 hours. I Cell viability (trypan blue assays) of A375 and SK-MEL-28 after silencing IDH1 compared to control (si.CTRL) under high and low glucose conditions for the indicated time points. Each data point represents the mean ± SEM of three independent experiments. N.S., nonsignificant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001