Extended Data Fig. 4: T cell-specific deletion of HK2 impairs T cell metabolism and immune responses against tumorigenesis and bacterial infection.
From: NF-κB-inducing kinase maintains T cell metabolic fitness in antitumor immunity
a,b, Genotyping PCR (a) and immunoblot (b) analysis of the Hk2tKO (tKO) and wildtype (WT) control mice. c,d, Seahorse analysis of basal ECAR (measured after glucose injection, Glc) and maximum ECAR (measured after oligomycin injection, Oligo) (c) and Seahorse analysis of baseline OCR (no treatment) and maximum OCR (FCCP injection) (d) in Hk2tKO or wildtype OT-I CD8 T cells either naïve or activated with anti-CD3 and anti-CD28 for 24 h. e, Flow cytometric analysis of the expression level of PD1 and Tim3 in CD8 T cells isolated from day-20 tumor of the B16F10-implanted Hk2tKO and wildtype control mice (n = 4 per genotype). f,g, Flow cytometric analysis of the frequency and absolute number of IFNγ-producing CD8 effector T cells in splenocytes (f) and bacterial burden in the spleen (g, presented as a representative image and a summary graph based on multiple mice) of Hk2tKO or wildtype mice infected i.v. with L. monocytogenes (1 × 105 CFU/mouse) for 7 (f) or 4 (g) days (f, n = 4 per genotype; g, n = 6 per genotype). h, Flow cytometric analysis of IFNγ-producing CD8 T cells in the spleen of the indicated mouse strains infected for 84 days with LCMV clone 13(4 × 106 PFU/mouse), restimulated in vitro with 3 μg/ml LCMV gp33-41 peptide for 14 h with monensin added during the last hour; WT(n = 6), Map3k14tKO(n = 6) and Hk2tKO(n = 3). Data are representative of two (f-h) or three (a-e) independent experiments. Summary data are shown as mean ± s.e.m. with P values determined by two-tailed Student’s t test.
