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. 2022 May 13;219(6):e20201405. doi: 10.1084/jem.20201405

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Figure S4. — Mouse immunophenotyping. (A) Whole blood was analyzed from 4-mo-old males and females using either the Pentraes 60 or Sysmex instruments and measured for hemoglobin content (HGB). n_{KO Female} = 5; n_{Control Female} = 7; n_{Control Male} = 4; n_{KO male} = 8. One of three experiments is shown. *, P < 0.05; **, P < 0.01. P values were calculated with an unpaired t-test. (B) Total splenocyte numbers of Gimap6^−/− and control mouse. n_KO = 19; n_Control = 23. Three experiments were pooled. ***, P < 0.001. P values were calculated with an unpaired t-test. (C) Representative flow plots of B (B220⁺), NK (NK1.1⁺), T (CD3⁺), and NKT (NK1.1⁺/CD3⁺) splenocyte fractions from KO and control mice. n_KO = 3; n_Control = 6. One of three experiments is shown. (D and E) Quantitation of C for cell fraction (D) or for absolute cell count (E). Three experiments were pooled. (D) n_KO = 7; n_Control = 9. (E) n_KO = 14; n_Control = 14. **, P < 0.01; ***, P < 0.001. Mann-Whitney U test. (F) Flow cytometry dot plots of naive (T_N; CD62L⁺CD44⁻), T_CM (CD62L⁺CD44⁺), T_EM (CD62L⁻CD44⁺), and effector (T_EFF; CD62L⁻CD44⁺) CD4⁺ and CD8⁺ T cells. One of three experiments is shown. (G) Quantification of naive and memory populations in F. n_KO = 3; n_Control = 6. ***, P < 0.001. P values were calculated with an unpaired t-test. (H) CXCR5 and PD-1 expression in CD4⁺ T cells. One of three experiments is shown. (I) Quantification of H. n_KO = 7; n_Control = 5. *, P < 0.05; **, P < 0.01. Mann-Whitney U test. (J) PD-1 expression in CD8 T cells. One of three experiments is shown. (K) Quantification of J. n_KO = 7; n_Control = 5. *, P < 0.05. Mann-Whitney U test. (L and M) Total serum IgM (L) and total serum IgG (M) were analyzed by ELISA. n_KO = 6; n_Control = 11. (N) Mice were injected intraperitoneally with 50 µg NP-CGG in PBS 1:1 with Imject alum adjuvant and boosted 35 d later. PBS with alum was a negative control. Serum was analyzed for NP-specific IgM and IgG₁ antibodies using an ELISA. Graphs show mean ± SD. AU, arbitrary units; NP, 4-Hydroxy-3-nitrophenylacetyl; NP-CGG, NP-chicken gamma globulin. n_PBS = 3; n_KO = 4; n_Control = 8. One of two experiments is shown. Bars (A, B, D, E, G, I, K, and L–N) represent mean ± SD.