Fig. 4: Impaired functionality of the thymic stroma in OVOL2 deficient thymi.
From: OVOL2 sustains postnatal thymic epithelial cell identity
A, B Real time-PCR analysis of Ovol2 transcripts from FACS sorted Ovol2+/+ cTECs and mTECs at E13.5 days, 0 days, 4 weeks, 12 weeks, 20 weeks, and 30 weeks of age (n = 3 per group). Mean and SD are shown in (A). The data are normalized to mean expression at E13.5 days (A) or to mean expression in mTECs (B). C, H, M Numbers of indicated subpopulations of thymic stromal cells in 4-wk-old (C), or 12-wk-old Ovol2C120Y/− mice (H), or 20-wk-old Foxn1-Cre;Ovol2fl/fl mice (M) and WT or Ovol2fl/fl littermates (n = 4 C120Y/−, 3 WT littermates in (C); 4 mice/genotype in (H); 3 mice/genotype in (M). Gating strategy is shown in Supplementary Fig. 7. D, I, N Quantitative analysis of early T-cell progenitors (ETP) in 4-wk-old (D), or 12-wk-old Ovol2C120Y/− thymi (I), or 20-wk-old Foxn1-Cre;Ovol2fl/fl thymi (N) and WT thymi (n = 3 C120Y/−, 4 WT littermates in (D); 4 mice/genotype in (I); 3 mice/genotype in (N). E, F, J, K, O, P Real time-PCR analysis of the indicated genes in FACS sorted mTECs (E, J, O) and endothelial cells (F, K, P) from 4-wk-old (E, F), or 12-wk-old Ovol2C120Y/− thymi (J, K), or 20-wk-old Foxn1-Cre;Ovol2fl/fl thymi (O, P) and WT thymi (n = 3 per group). G, L Real time-PCR analysis of the indicated genes in FACS sorted cTECs from 4-wk-old (G), or 12-wk-old Ovol2C120Y/− thymi (L) and WT littermates (n = 3 per group). Q, R Surface marker MHC-II (Q) and MHC-I (R) expression by cTECs in 4- or 12-wk-old Ovol2C120Y/− mice and WT littermates (n = 3 mice/genotype). S Real time-PCR analysis of Aire and Fezf2 from FACS sorted Ovol2+/+ and Ovol2C120Y/C120Y mTECs at 4.5 weeks of age (n = 3 per group). T Intracellular AIRE intensity in total or MHC-IIhigh mTECs of 4.5-wk-old Ovol2C120Y/C120Y and WT littermates (n = 5 C120Y/C120Y, 6 WT littermates). U, V Surface marker CD80 (U) and CD86 (V) expression by total or MHC-IIhigh mTECs in 4.5-wk-old Ovol2C120Y/C120Y mice and WT littermates (n = 8 mice/genotype). Data are representative of two (A–R, T–V) or four independent experiments (S). Data points represent individual mice (C, D, H, I, M, N, Q, R, T–V) or individual sorting pools from 5 mice per pool (A, B, E–G, J–L, O, P, S). Means ± SD are indicated and P values were determined by two-tailed Student’s t test. MFI, mean fluorescence intensity. +/+, WT; C120Y/−, Ovol2C120Y/−; C120Y/C102Y, Ovol2C102Y/C120Y; and fl/fl, Ovol2fl/fl.
