Extended Data Figure 1: Characterization of Fh1-deficient and Fh1-rescued cells.

a, PCR to assess Fh1 recombination. Putative genotypes are indicated on the right and are based on the expected size of the genomic PCR amplification products as from ref. 7. Fh1^fl/fl = 470 bp and Fh1^−/− = 380 bp. b, Fh1 protein levels measured by western blotting of cells of the indicated genotype. Calnexin was used as loading control. c, Intracellular fumarate levels measured by LC–MS and normalized to total ion count. Results were obtained from four independent cultures and are indicated as average ± s.d. P values were calculated from one-way ANOVA. d, OCR and ECAR assessed using the Seahorse Extracellular Flux Analyser. Results were obtained from five replicate wells and are presented as average ± s.d. e, Bright-field images of cells of the indicated phenotype. Scale bar, 400 μm. Western blot and gel source images are presented in Supplementary Fig. 1. Source data are presented in Supplementary Table 2. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. f, Schematic representation of the proposed link between loss of FH, fumarate accumulation, and epigenetic suppression of the antimetastatic cluster of miRNA miR-200. Upon accumulation of fumarate as a result of FH inactivation, the TET-mediated demethylation of the mir-200ba429 cluster is inhibited, leading to their epigenetic suppression. As a consequence, Zeb1 and Zeb2 are de-repressed, eliciting a signalling cascade that leads to EMT.