Extended Data Fig. 8: Inhibition of IDH2 induces metabolic compensation.

a, MitoSOX MFI in DMSO- or IDH2i-treated cells upon addition of H₂O₂ at indicated concentrations, measured by flow cytometry. (n = 5 biological replicates, pooled data from 2 independent experiments). b, Number of live TFAM WT and TFAM KO T cells per millilitre upon DMSO or IDH2i treatment. (n = 3 biological replicates, pooled data from 2 independent experiments). c, Percentage of CD62L⁺ out of CD8⁺ TFAM WT and KO T cells upon DMSO or IDH2i treatment. (n = 3 biological replicates, pooled data from 2 independent experiments). d, Percentage of CD62L⁺ out of Thy1.1⁺ CD8⁺ T cells at day 7 post-activation upon deletion of indicated genes. (n = 6 (Scr, Ogdh and Sdhb) and n = 5 (Idh2) biological replicates per group, pooled data from 3 independent experiments). e, Immunoblots for OGDH and SDHB in control Scr and OGDH- or SDHB-deleted cells. (Representative of 2 independent experiments). f, Percentages of citrate m + 5 detected by [U-¹³C]-glutamine labelling in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). g, Percentages of m + 3 succinate, fumarate and malate detected by 2 h [U-¹³C]-glutamine labelling in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). h, Percentages of m + 4 succinate, fumarate and malate detected by 2 h [U-¹³C]-glutamine labelling in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). i, Percentages of m + 5 α-KG detected by 2 h [U-¹³C]-glutamine labelling in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). j, Percentages of m + 2 acetyl-CoA, detected by [U-¹³C]-glucose labelling for indicated time in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). k, Percentages of m + 2 citrate, succinate, fumarate and malate detected by 2 h [U-¹³C]-glucose labelling in DMSO or IDH2i-treated T cells. (n = 3 biological replicates per group). l, Percentages of citrate m + 2 detected by [U-¹³C₁₆]-palmitate labelling in mouse CD8⁺ T cells treated with DMSO or the IDH2 inhibitor (IDH2i). (n = 3 biological replicates per group). m–o, Quantification of basal OCR (m), ATP-linked OCR (n) and spare respiratory capacity (SRC; o) from data presented in Fig. 4d. (n = 4 biological replicates, pooled data from 2 independent experiments). p, Percentage of CD62L⁺ T cells in control Scr or ΟGDH-deleted CD8⁺ T cells, treated with IDH2i. (n = 4 biological replicates, pooled data from 2 independent experiments). q, Percentage of CD62L⁺ T cells in DMSO or IDH2i-conditioned CD8⁺ T cells, treated with etomoxir (FAOi). (n = 5 biological replicates, pooled data from 2 independent experiments). r, Glucose consumption in DMSO- or IDH2i-treated mouse CD8⁺ T cells over 24 h, at day 3 post-activation. (n = 3 biological replicates per group). s, Intracellular abundances of indicated amino acids (arbitrary units, a.u) measured by mass spectrometry upon DMSO or IDH2i treatment. (n = 9 biological replicates, pooled data from 3 independent experiments). t,u, CD98 MFI in DMSO- or AG-221- (t) or AGI-6780- (u) treated CD8⁺ T cells, measured by flow cytometry at day 3 post-activation. (n = 9 (AG-221) and n = 3 (AGI-6780) biological replicates, pooled data from 4 (AG-221) and 2 (AGI-6780) independent experiments). Data represent mean ± s.e.m. and were analysed by unpaired two-tailed Student’s t-test (f,i,l–o,r,t,u), multiple unpaired, two-tailed t-tests using Benjamini and Hochberg method (g,h,j,k,s), one-way ANOVA using Tukey’s multiple comparison test (d) or two-way ANOVA using the original FDR test of Benjamini and Hochberg (a–c,p,q). Only relevant statistical comparisons are shown. For gel source data, see Supplementary Fig. 1.

Source data