Mitochondrial <span>l</span>-2-hydroxyglutarate is a physiological signalling metabolite

Cell culture, drug treatment and fractionation 143B cells were cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, 11965-126) supplemented with 10% Nu-serum IV (Corning, 355504), 10 mM HEPES (Corning, 25-060-CI), 1× GlutaMAX (Gibco, 35050061), 1× antibiotic–antimycotic solution (Gibco, 15240062), 2.5 µg ml −1 plasmocin prophylactic (InvivoGen, ant-mpp), 1 mM methyl pyruvate (Sigma-Aldrich, 371173) and 400 µM uridine (Sigma-Aldrich, U3003). Mouse ES cells were cultured in 2i medium consisting of 0.5× Neurobasal medium (Gibco, 21103049), 0.5× DMEM/F12 (Gibco, 11320033), 0.5× N-2 supplement (Gibco, 17502048), 0.5× B-27 supplement (Gibco, 17504-044), 0.05% bovine albumin fraction V (Gibco, 15260037), 1× antibiotic–antimycotic solution (Gibco, 15240062), 2.5 µg ml −1 plasmocin prophylactic (InvivoGen, ant-mpp), 1 μM PD0325901 (Reprocell, 04-0006-02), 3 μM CHIR99021 (Reprocell, 04-0004-02), 2 mM glutamine (Gibco, 25030081), 150 μM 1-thioglycerol (Sigma-Aldrich, M6145), 1,000 U ml –1 leukaemia inhibitory factor (Sigma-Aldrich, ESG1106), 1 mM methyl pyruvate (Sigma-Aldrich, 371173) and 400 µM uridine (Sigma-Aldrich, U3003). …