Fig. 7: Eed^fl/fl::ZsGreen::β5tCre mice contain a subpopulation of Eed-proficient mTEC.

a ZsGreen expression in cTEC and mTEC isolated from 4-week-old Eed^fl/fl::ZsGreen::β5tCre mice. b Quantification of H3K27me3 abundance in ZsGreen-positive and -negative TEC and their phenotypic characterization. c Representative histograms of histone H3 abundance in ZsGreen-positive and -negative TEC. d Representative histograms and median fluorescence quantification of H3K27me2 and EZH2. Fluorescence of control mTEC^hi was set as 100%. Data are pooled from two independent experiments identified by circles and diamonds, respectively. e Representative FACS plots and quantitative analysis of cTEC, mTEC and mTEC^hi frequencies within ZsGreen-positive and -negative TEC. f, g Representative histograms and quantitative analysis of ZsGreen-positive and -negative cTEC (f) and mTEC^hi (g) phenotypes. h Spatial distribution of ZsGreen-positive and -negative TEC in thymus tissue sections of 4-week-old Eed^fl/fl::ZsGreen::β5tCre mice. Immunofluorescence analysis of mTEC expressing ZsGreen and cytokeratin 14 (CK14) in the medulla (marked by dashed blue line) identifying single cells and small aggregates of ZsGreen-negative cells (white dashed lines). Data in bar graphs show mean ± SD and are pooled from two independent experiments (a, b, d) or are representative of two independent experiments (e–g). Contour plots and histograms are representative for data displayed as frequencies in bar graphs (a, b, d, e–g). Immunofluorescence data are representative of two independent experiments with 3 sections each. a, b Eed^fl/fl::ZsGreen::β5tCre (n = 3), in (b) dark green represents ZsGreen-positive H3K27me3-positive, light green shows ZsGreen-positive H3K27me3-negative, and grey depicts ZsGreen-negative H3K27me3-positive, (d) Eed^fl/fl::ZsGreen (n = 5, grey histograms and bars with black circles and diamonds) Eed^fl/fl::ZsGreen::β5tCre (n = 7, ZsGreen-negative: white histograms and bars with grey circles and diamonds, ZsGreen-positive: black histograms and bars with grey circles and diamonds), (e–g) Eed^fl/fl::ZsGreen (n = 5, grey histograms and bars with black circles), Eed^fl/fl::ZsGreen::β5tCre (n = 5, ZsGreen-negative: white histograms and bars with grey diamonds, ZsGreen-positive: black histograms and bars with grey diamonds); n = biologically independent replicates per group, mice were 3–4 weeks of age. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 (two-tailed unpaired Student’s t test). Source data including exact statistical test values are provided in the Source Data file.