Figure 1. T cell-specific NIK ablation does not affect thymocyte development.

(a) Genotyping PCR analysis of wildtype (WT) and floxed alleles of NIK as well as Cre using tail DNA of the indicated mice. (b) Immunoblotting analysis of NIK using whole cell lysates of WT and NIK-TKO naïve CD4⁺ T cells treated with 10 μM MG132 for 5 h (to enhance NIK level). (c) Immunoblotting analysis of p100 and p52 using whole cell lysates from naïve CD4⁺ T cells stimulated for 48 hours with plate-bound anti-CD3 plus anti-CD28. (d) Flow cytometry analyses of B cells (B220⁺CD3⁻) in the spleen of age-matched 6–8 weeks old WT and NIK-TKO mice. Data are representative of 3 independent experiments. (e,f) Flow cytometry analyses of thymocyte and T cell populations in the thymus and spleen of age-matched (6–8 weeks old) WT and NIK-TKO mice based on CD4 and CD8 staining. Data are presented as a representative plot (e) and mean ± SD values of multiple mice (f, each circle represents a mouse). (g,h) Flow cytometry analysis of the surface expression of CD69 and TCRβ on thymocytes from 5 weeks old WT and NIK-TKO mice, showing four subpopulations (1, TCR^loCD69^lo; 2, TCR^loCD69^hi; 3, TCR^hiCD69^hi; 4, TCR^hiCD69^lo) of thymocytes, presented as a representative plot (g) and and mean ± SD values of multiple mice (h).