Fig. 1: NK-specific gp96 deficiency reduces NK cell maturation.

A Western blot of gp96 expression in splenic NK cells sorted from WT and KO mice. Representative flow cytometry plots showing the percentages of NKp (NK1.1^-DX5^-), iNK (NK1.1⁺DX5^-), mNK (NK1.1⁺DX5⁺) cells gated on CD3^-CD122⁺ splenocytes (B) and bone marrow (C) cells and representative flow cytometry plots showing the percentages of CD27^-CD11b^- (DN), CD27⁺CD11b^- (CD27 SP), CD27⁺CD11b⁺ (DP), and CD27^-CD11b⁺ (CD11b SP) cells on gated NK1.1⁺DX5⁺ splenocytes (B) and bone marrow (C) cells from WT and gp96-deficient mice. The numbers are percentages of the indicated quadrants among the gated cells. D Flow cytometry analysis of indicated marker levels. E Expression of indicated markers as determined by RNA-seq. The fold change indicates the difference in relative transcript expression between WT compared with Ncr1^Cregp96^fl/fl mice. F–H The chimeric mouse model was produced as in (F). Flow cytometry analysis of CD27 and CD11b on DX5⁺ NK cells from CD45.1⁺ cells and CD45.2⁺ cells in the spleen (G) and bone marrow (H). The data are representative of two independent experiments with similar results. Dots represent data from n = 5 mice/group. Mean ± SD is shown. Statistical significance was determined using two-tailed unpaired t test. *p <  0.05, **p <  0.01, ***p <  0.001, ****p <  0.0001. p values: (B) p = 0.002 (NKp), p < 0.0001 (iNK), p < 0.0001 (mNK), p < 0.0001 (DN), p < 0.0001 (CD27 SP), p < 0.0001 (DP), p < 0.0001 (CD11b SP), (C) p = 0.009 (NKp), p < 0.0001 (iNK), p < 0.0001 (mNK), p = 0.0427 (DN), p < 0.0001 (CD27 SP), p < 0.0001 (DP), p < 0.0001 (CD11b SP), (G) p < 0.0001 (NKp), p < 0.0001 (iNK), p < 0.0001 (mNK), p < 0.0001 (DN), p = 0.0011 (CD27 SP), p = 0.0002 (DP), p < 0.0001 (CD11b SP), (H) p = 0.0281 (NKp), p < 0.0001 (iNK), p < 0.0001 (mNK), p < 0.0001 (DN), p = 0.0002 (CD27 SP), p = 0.0008 (DP), p < 0.0001 (CD11b SP).