Extended Data Fig. 1: ABHD11 deficiency leads to the accumulation of glutaryl-lipoate adducts.
From: Lipoyl deglutarylation by ABHD11 regulates mitochondrial and T cell metabolism
a, Lp or Kglu immunoblot in HeLa cells treated with ML226 or DEG for the indicated times. Representative of n = 2 independent experiments. b, HeLa cells were treated with 1 µM ML226 for 24 h, or Cas9-expressing HeLa cells were transduced with sgRNAs targeting β2M (control) or ABHD11 for 11 days. DLST (OGDHc-E2) mRNA levels were determined using qRT-PCR. Transcript levels are normalized to β-actin (ACTB) and adjusted relative to the control condition. Mean ± s.d.; n = 4 independent experiments; one-way ANOVA and Dunnett’s post hoc test. c, Cas9-expressing HeLa cells were transduced as in (b) and treated with 10 µg/ml cycloheximide (CHX) with or without 2.5 µM ML226 for the indicated times. Levels of OGDHc-E2 were determined using immunoblotting, with HIF1α as a control. Representative of n = 4 independent experiments. d, OGDHc-E2 immunoprecipitation in HeLa cells for LC–MS/MS analysis of Lp modifications in Fig. 1h–k. Representative of n = 4 independent experiments. e–g, Representative chromatograms of OGDHc-E2 K110 Lp (e) in Fig. 1h; K110 Lpglu (f) in Fig. 1i; and K110 Lpglu (g) in Fig. 1j.
