Figure 1. RANKL is upregulated in CD4⁺ SP and LTi cells during the course of thymic regeneration.

• A
  Expression of RANKL protein analyzed by flow cytometry in CD45⁻ and CD45⁺ thymic cells from untreated (UT) WT mice or at d3 SL‐TBI.
• B, C
  Flow cytometry profiles and frequencies of DN (double negative), DP (double positive), CD4⁺ and CD8⁺ SP (single positive) (B), and LTi cells (C) from untreated (UT) WT mice or at d3 SL‐TBI.
• D
  Expression level of RANKL protein in CD4⁺ SP and LTi cells from UT WT mice or at d3 SL‐TBI and L‐TBI.
• E
  Expression of Rankl mRNA in the total thymus isolated from UT WT, Rorc^−/−, ZAP‐70^−/−, and Rag2^−/− mice or at d3 SL‐TBI (n = 3–6 mice per genotype).
• F
  CD4⁺ SP and LTi cells from UT WT mice or at d3, d6, d10, and d20 SL‐TBI with no hematopoietic rescue were analyzed for the expression of RANKL protein. Mean fluorescence intensity (MFI) of RANKL in CD4⁺ SP and LTi cells over time following SL‐TBI. The red lines represent the MFI of RANKL at baseline.

Data information: Data are shown as mean ± SEM and are pooled of four independent experiments with similar results (n = 3–4 mice per group). *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001. Exact P‐values and statistical tests used to calculate them are provided in Appendix Table S2.