Fig. 2

Knock-down of ERK1/2 by siRNAs and treatment with a MEK1 inhibitor induces pS₃₀₀PDH. a A375 cells were transfected with siRNA against ERK1/2 (50 nM each siRNA) or a scrambled control (100 nM) for 72 h. 48 h prior to collection, the cells were incubated with either PLX4032 (1 μM) or Trametinib (3 nM) or both drugs. One representative of three biological replicates is shown. b Western blot analysis of A375 (BRAF^V600E), MelJuso, IPC298 and SKMel30 cells (NRAS^mut) treated with 5 nM of Trametinib for the indicated time points. α-Tubulin was used as loading control; representative blots of three biological replicates are shown. 1: Untransfected; 2: scrambled; 3: ERK1/2 siRNA