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. 2020 Dec 29;218(4):e20201311. doi: 10.1084/jem.20201311

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© 2020 Michaels et al.

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Figure S3. — Analysis of cell-intrinsic effects of Nr1h4 deficiency on T cell differentiation in the thymus. CD45.2⁺ BM precursor cells from CD4-CreNr1h2^+/+, CD4-CreNr1h2^fl/+, or CD4-CreNr1h2^fl/fl animals were mixed at a 1:1 ratio with CD45.1⁺ calibrator cells from WT donors and transferred into lethally irradiated mice. Thymi of recipient mice were analyzed 8 wk after reconstitution. (A) Representative flow cytometry plots comparing CD4-CreNr1h2^+/+ and CD4-CreNr1h2^fl/fl T cell development. (B) Contribution of CD45.2⁺ cells with the indicated genotype to the DN, DP, CD4SP, and CD8SP populations. (C) Frequencies of Annexin V⁺ cells among CD45.2⁺ CD4SP and CD8SP thymocyte subsets. (D) Frequencies of Foxp3⁺ cells among CD45.2⁺CD4SP thymocytes. (E and F) Representative flow cytometry plot (E) and quantification (F) of T reg cell precursor subsets among CD45.2⁺ CD4SP cells. Data are presented as mean ± SEM (n = 4–7). ns, nonsignificant = P > 0.05; *, P < 0.05; **, P < 0.01; ****, P < 0.0001. Statistical significance was determined using one-way ANOVA followed by the Holm-Šídák correction. Data are representative of three independent experiments.