FIGURE 2.

Extent of 2HG accumulation correlates with the degree of differentiation blockade in non-transformed cells expressing IDH2 Arg-140 or Arg-172 mutations. A, Western blot of 3T3-L1 pre-adipocytes stably expressing additional IDH2 WT, IDH2 R172K, IDH2 R140Q, or empty vector. IDH1 protein levels were used as a loading control. B, cells were treated with a mixture to induce differentiation into mature adipocytes for 7 days. Oil Red O staining was used to assess the accumulation of lipid droplets. C, at day 4 following differentiation induction, RNA was extracted and the expression of adipocyte-specific gene and transcription factors was measured by quantitative PCR with reverse transcription (RT-qPCR). B and C, data are representative of 3 independent experiments. D, cellular 2HG accumulation was measured by GC-MS. Data are representative of mean ± S.D. of 3 biological replicates from 2 independent experiments. *, p < 0.001.