Extended Data Fig. 10: AT2 mTECs expand in juvenile Foxn1-Cre(+)/Runx1fl/fl/CBG mice and can develop through an Aire-expressing stage or independently of Aire.
From: Thymic alveolar type 2 epithelial mimetic cells revealed by RUNX1 deficiency
a-c, Flow cytometry and percentage of GFP+ mTECs from Foxn1-Cre(+)/Runx1fl/fl/CBG mice (Cre(+)/CBG) at the indicated timepoints (n = 2 (Day 1), n = 2 (Day 4), n = 4 (Day 7), n = 4 (Day 30), and n = 2 (Day 140); two independent experiments). d-f, Flow cytometry and percentage of MHC-IIhigh and MHC-Illow mTECs in Foxn1-Cre(-)/Runx1fl/fl/CBG mice (Cre(-)/CBG) mTECs and Foxn1-Cre(+)/Runx1fl/fl/CBG mice (Cre(+)/CBG) mice at the indicated timepoints. g, IGV CUT&RUN tracks for the indicated genes. h, Cre(-)/CBG mice were treated with EGF for either two or three weeks and flow cytometry was performed for GFPlow and GFPhigh AT2 mTEC (n = 5 mice treated with vehicle for 2 weeks, n = 5 mice treated with EGF for 2 weeks, n = 2 mice treated with vehicle for 3 weeks, and n = 3 mice treated with EGF for 3 weeks; two independent experiments). i, Flow cytometry (left) and percentage and absolute number (right) of GFP+ mTECs from 30-day-old Cre(-)/CBG and Foxn1-Cre(+)/Runx1+/fl/CBG mice (Het/CBG) 21 days after irradiation and treated with EGF or Vehicle for three weeks (n = 1 vehicle-treated Cre(-)/CBG, n = 2 EGF-treated Cre(-)/CBG, n = 1 vehicle-treated Het/CBG, and n = 1 EGF-treated Het/CBG mice; representative experiment). j, Cre(-)/CBG mice were treated with EGF for three weeks and qPCR of AT2 marker genes was performed on sort purified GFPlow, GFPhigh, and a control population (GFPneg) (n = 3 mice; representative experiment). k, qPCR of Runx1 gene expression from sorted MHC-IIlow mTECs from Foxn1-Cre(-)/Runx1fl/fl mice (Cre(-)/Runx1fl/fl), Foxn1-Cre(+)/Runx1+/fl mice (Cre(+)/Runx1+/fl), and Foxn1-Cre(+)/Runx1fl/fl mice (Cre(+)/Runx1fl/fl) (n = 4 Cre(-)/Runx1fl/fl, n = 2 Cre(+)/Runx1+/fl, and n = 2 Cre(+)/Runx1fl/fl mice; representative experiment). l-m, Flow cytometric analysis of CD11c−/CD45−/EpCAM+/Ly51−/tdtTmt+/GFP+ mTECs from (l) Aire-CreERT2/R26tdTomato/ Runx1+/fl (CBG(-)) mice or (m) Aire-CreERT2/R26tdTomato/Runx1+/fl/ CBG (CBG(+)) mice treated with tamoxifen and EGF for two weeks (lineage trace). n, Flow cytometry (left) and percentage and absolute number (right) of GFP+ mTECs from 30-day-old Het/CBG mice treated with IGF2 or Vehicle for two weeks (n = 2 mice; representative experiment). (h,j) In graphs, the bars correspond to the mean, with error bars showing ±SD of values shown, and each data point represents an individual mouse. (h,j) Statistical significance was calculated using a one-way ANOVA, and grouped comparisons were corrected using Tukey’s multiple comparison test.
