Extended Data Fig. 4: Haem drives terminal differentiation of CD8+ T cells via the BACH2-BLIMP1 axis.
From: Proteasome-guided haem signalling axis contributes to T cell exhaustion
a-d, Representative flow plots (a, c) and quantitative results (b, d) of BACH2 and BLIMP1 in MDR/MGlow and MDR/MGhigh subsets under treatment with Oligomycin A, Antimycin A, and Mdivi-1 (BACH2, n = 4; BLIMP1, n = 3). e-h, BACH2 (e, f) and BLIMP1 (g, h) expression in TPEx and TTEx P14 TILs (BACH2, n = 20; BLIMP1, n = 19). i, Regulatory haem concentration in normal FBS and haem-depleted FBS (n = 3). j, Luminescence of the indicated Jurkat reporter cells cultured in normal or haem-depleted FBS under PMA and Ionomycin stimulation (n = 3). k, Western blot plots of HA-tagged BACH2 and β-actin in the indicated Jurkat reporter cells cultured in normal or haem-depleted FBS. HA-tagged BACH2 and β-actin were run on the same gel as loading controls. For gel source data, see Supplementary Fig. 1. l, Representative flow plots show BLIMP1 in Bach2WT or Bach2MUT overexpressed- Bach2CKO P14 cells treated with or without haemin under repetitive stimulation. m,n, Representative flow plots of TCF-1+PD-1+TIM3− cells (TPEx), TCF-1+PD-1+TIM3+ cells (TIEx), and TCF-1−PD-1+TIM3+ cells (TTEx) in tumours (m) and TCF-1+TOX− cells (TTSM) and TCF-1+TOX+ cells (TPEx) in dLN (n). o, Representative flow plots of BLIMP1 in Bach2WT or Bach2MUT P14 cells in tumours and dLNs. p, Representative flow plots of MDR and MG staining in Bach2WT or Bach2MUT P14 TILs. q, Average expression of subset markers across P14 TIL clusters and ProjecTILs reference datasets used for annotation. Data are representative of two (j, k) or three (i) independent experiments with similar results or are the cumulative results from three (b, d, f, h) independent experiments. Each symbol represents one individual. Data are means ± s.d. and were analysed by two-tailed, unpaired student’s t test (b, d, f, h, i) or two-way ANOVA (j). TIL, tumour-infiltrating lymphocyte; dLN, draining lymph node; CKO, conditional knockout; TPEx, Progenitor exhausted T; TIEx, Intermediate exhausted T; TTEx, Terminal exhausted T; TTSM, tumour specific memory T.
