Extended Data Fig. 1. NIK regulates T cell exhaustion and antitumor immunity.

a, genotyping PCR of R26Stop^FLMap3k14 (Map3k14^Tg), Map3k14^+/+CreER, and R26Stop^FLMap3k14-CreER (Map3k14^TgCreER) mice, showing the PCR products of NIK-GFP in Map3k14^Tg allele and CreER. b, Immunoblot analysis of NIK expression in tamoxifen-treated Map3k14^+/+CreER (WT) and Map3k14^TgCreER (iTg) mice. c, Schematic of experimental design for producing B16F10 tumor-bearing NIK^iTg and WT control mice. Each mouse was injected s.c. with 5 x 10⁵ B16F10 cells (WT=9, iTg=7). d,e, Flow cytometric analysis of the frequency and absolute cell number of CD4 and CD8 T cells in the tumor (d) or draining lymph node (e) of day 18 B16F10 tumor-implanted NIK-iTg and WT mice (d, WT: n=4; iTg: n=5; e, n=6 per genotype). f,g, Flow cytometric analysis of the frequency and absolute number of CD44⁺CXCR3⁺ CD8 effector T cells in the draining lymph node (f) or tumor (g) of day 18 B16F10 tumor-implanted NIK-iTg and wildtype mice (f, n=6 per genotype; g, n=4 per genotype). h,i, Flow cytometric analysis of the frequency and absolute number of PD1⁺Tim3⁺ CD8⁺ T cells (h) or IFNγ-producing PD1⁺Tim3⁺ CD8 T cells (i) in the tumor of day 18 B16F10-implanted NIK^iTg and wildtype control mice (h,i, n=4 per genotype). Data are representative of three independent experiments. Summary data are shown as mean ± s.e.m. with P values determined by two-tailed Student’s t test.