Extended Data Fig. 4: Loss of U34 enzymes impairs HIF1α-dependent metabolism.
From: Codon-specific translation reprogramming promotes resistance to targeted therapy
a, b, MM117 (BRAFWT) primary line was depleted of ELP3 (a) or CTU2 (b). Lactate and 13C-glucose-derived lactate produced in the medium was quantified by NMR over time. n = 3 independent experiments, two-sided t-test, data are mean + s.d. c, d, MM029 (BRAFV600E) primary line was depleted of ELP3 (c) or CTU2 (d). Lactate and 13C-glucose derived lactate produced in the medium was quantified by NMR over time. n = 3 independent experiments, two-sided t-test, data are mean + s.d. e–g, Human short-term primary melanoma lines were depleted of HIF1A by esiRNA. e, Indicated proteins were detected by western blot (e, n = 1 independent experiment). f, Death of cells in e was quantified by FACS. n = 2 independent experiments, data are mean + s.d. g, Indicated mRNA levels of cells in e were quantified by qPCR. n = 2 independent experiments, data are mean + s.d. h, i, B16 melanoma cells stably expressing pBABE-control or pBABE-BRAF(V600E) were depleted of Hif1a by esiRNA. h, Indicated protein levels were detected by western blot. n = 1 independent experiment. i, Cell death was quantified by FACS. n = 2 independent experiments, data are mean + s.d. j–l, A2058 cells were depleted of HIF1A by esiRNA. j, Indicated protein levels were detected by western blot. n = 1 independent experiment. k, Cell death was quantified by FACS. n = 2 independent experiments, data are mean + s.d. l, The expression of the indicated mRNAs was detected by qPCR. n = 2 independent experiments, data are mean + s.d.
