Fig 1: BMI1 induces NLR family CARD domain containing 5 (NLRC5) degradation and promotes immune escape in NSCLC. (A) protein levels of BMI1, NLRC5, and human leukocyte antigen class I in A549 and H1299 cells determined by immunoblot analysis (n = 3); (B) expression of CD25 in CD8+ T cells co‐cultured with NSCLC cells examined by flow cytometry (n = 3); (C) IL‐2 production in CD8+ T cells co‐cultured with NSCLC cells examined using ELISA kits (n = 3); (D) PD‐1 expression in CD8+ T cells after co‐culture with A549 and H1299 cells examined by flow cytometry (n = 3); (E and F) mRNA (E) and protein (F) levels of PD‐L1 in A549 and H1299 cells co‐cultured with CD8+ T cells examined by reverse transcription‐quantitative polymerase chain reaction and immunoblot assays (n = 3). Measurement data are expressed as the mean ± SD. Differences are analyzed by the one‐way ANOVA (C, E, and F) or two‐way ANOVA (A and B). *p < 0.05 versus the sh‐NC group; # p < 0.05 versus the oe‐NC + oe‐NLRC5 group
Fig 2: BMI1 binds to NLR family CARD domain containing 5 (NLRC5) to promote its ubiquitination and protein degradation. The NLRC5 degradation leads to human leukocyte antigen class I downregulation, which suppresses the cytotoxicity of activated CD8+ T cells and production of IL‐2 to promote immune evasion in NSCLC
Fig 3: NLR family CARD domain containing 5 (NLRC5) upregulates human leukocyte antigen (HLA) class I expression, promotes T cell activation and IL‐2 production, and suppresses PD‐1/PD‐L1 levels. (A) NLRC5 expression in A549 and H1299 cells after oe‐NLRC5 transfection determined by reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) (n = 3); (B) mRNA (B) and protein (C) levels of HLA class I in A549 and H1299 cells after NLRC5 overexpression determined by RT‐qPCR and immunoblot analysis (n = 3); (D) HLA class I expression in A549 and H1299 cells after NLRC5 overexpression evaluated by flow cytometry (n = 3); (E) expression of CD25 in the activated CD8+ T cells after co‐cultured with A549 and H1299 cells analyzed by flow cytometry (n = 3); (F) IL‐2 production in the activated CD8+ T cells after co‐culture with A549 and H1299 cells examined using ELISA kits (n = 3); (G) PD‐1 expression in the CD8+ T cells after co‐culture with A549 and H1299 cells examined by flow cytometry (n = 3); (H and I) mRNA (H) and protein (I) levels of PD‐L1 in A549 and H1299 cells after co‐culture with CD8+ T cells analyzed by RT‐qPCR and immunoblot assays (n = 3). Measurement data are expressed as the mean ± SD. Differences were analyzed by two‐way ANOVA (A–I). *p < 0.05
Supplier Page from Abcam for Human IL-2 ELISA Kit