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. 2020 Jul 27;217(10):e20200318. doi: 10.1084/jem.20200318

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© 2020 Chan et al.

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Figure 3. — LXRαβ-deficient TECs are functionally competent but display impaired proliferation. (A) Representative flow cytometry plots and quantification over time from live FoxN1-LXRαβ⁻^/− versus LXRαβ^fl/fl TECs (n = 5–8 per group from two independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001. (B–D) Experimental approach (B) and enumeration of engrafted GFP⁺CD45⁺ and GFP⁺CD45⁻ cells (C) and thymus weight and cellularity (D) of FoxN1-LXRαβ⁻^/− mice intrathymically injected GFP⁺CD45⁻ or GFP⁺CD45⁻ cells. Data are n = 4 or 5 per group from two independent experiments. *P < 0.05, **P < 0.01. (E) Histological staining of keratin-5 (K5) and keratin-8 (K8) from 2-mo-old Lxrαβ^fl/fl and FoxN1-Lxrαβ⁻^/− mice (representative of three mice). White boxes indicate zoomed-in section represented in the lower panels. (F) In 2-mo-old C57BL6/J (WT) or LXRαβ⁻^/− mice, Foxn1, Il7, Psmb11, and Aire levels by real-time qPCR from sorted CD45⁺, cTECs, and mTECs (n = 4–6 per group from two independent experiments). (G) Frequency of CD4⁺ SP or CD8⁺ SP thymocytes that express different TCRVβ chains (n = 3 or 4 per group, representative of two independent experiments). (H–J) From 2-mo-old Lxrαβ^fl/fl and FoxN1-Lxrαβ⁻^/− mice, Caspase (Casp) 3/7⁺ cTECs and mTECs (H; n = 5 or 6 per group from two independent experiments), representative flow cytometry data and quantification of BrdU⁺ (I; injected 24 h prior), and ki67⁺DAPI⁺ cTECs and mTECs (J). (I and J) Data are n = 9 per group from two or three independent experiments. **P < 0.01, ***P < 0.001. (K) Experimental approach (n = 4 or 5 per group). (L and M) Proportion of BrdU⁺ cTECs and mTECs after 4-wk BrdU delivery (L) and representative flow cytometry images and quantification of BrdU⁺ cTECs and mTECs after the 2-wk washout BrdU withdrawal period (M). (K–M) Data are four or five per group from two independent experiments. *P < 0.05, **P < 0.01. Statistical analysis used was Student’s t test or two-way ANOVA followed by Sidak’s post-test. All data are mean ± SEM. Sac, sacrifice; i.th., intrathymic injection.