Extended Data Fig. 6: NFX1 is involved in CARNS2 and CD8+ T-cell-mediated anti-tumor immunity.
From: Carnosine regulation of intracellular pH homeostasis promotes lysosome-dependent tumor immunoevasion
a-d, At the end of the experiment in Fig. 4a, tumors were collected and tumor masses were calculated (n = 5 mice) (a). Immunoblotting verified CARNS2, NFX1, galectin-9 expression in the extracted tumors (b). Representative flow staining (c) and frequency of exhaustion and effector function markers (d) of CD8+ T cells in NTC and shCARNS2 Hepa 1-6 tumors (n = 3 samples). e, Corresponding to the experiment in Fig. 4c, tumors were collected (n = 5 mice) (left). Immunoblotting verified CARNS2, NFX1 and galectin-9 expression in the extracted tumors (right). f, Plasmids expressing human YAP-5SA alone or human YAP-5SA plus shNFX1 plasmids together with PB transposase plasmids were delivered by hydrodynamic injection into WT or Carns1&2-/- mice. Liver/body weight ratios were measured approximately at 100 days after injection (n = 5 mice). CD8+ T cells numbers, exhaustion and effector function markers were determined by flow cytometry (n = 3 samples). g, Flow cytometry analysis of Ki67, TIM-3, TNFα, INFγ expression on in vitro differentiated CD8+ T cells cultured with CM from Hepa 1-6 cells expressing EV or NFX1. Hepa 1-6 cells were treated under 20% O2 or 1% O2 for 48 h (n = 3 biological replicates). Data presented as mean ± s.d.(a, d, f, g). Statistical significance was determined by two-tailed unpaired Student’s t-test (a, d, f, g).
