Fig. 3: OVOL2 function in TECs is necessary to support thymic T cell development.

A Representative photographs of thymi from 4- or 12-wk-old Ovol2^C120Y/− and WT littermates. B Representative Oil red O staining of Ovol2^C120Y/− and WT littermate thymi at 12 wk of age. C–I Representative flow cytometry plots of TECs (C–E, H) and quantitative analysis (F, G, I) of indicated subsets in the thymus (F, I) or TECs (G) from 12-wk-old (C–G) or 4-wk-old (H, I) Ovol2^C120Y/− mice and WT littermates. Numbers adjacent to outlined regions indicate percent cells in each (mean ± SD) (n = 3 C120Y/−, 4 WT littermates in (C–E); 4 mice/genotype in (F); 4 (epithelial, cTEC, mTEC, mTEC^AIRE(high)) or 3 (mTEC^MHC-II(high)) C120Y/− mice, 5 (cTEC, mTEC, mTEC^AIRE(high)) or 4 (epithelial, mTEC^MHC-II(high)) WT littermates in (G); 3 mice/genotype in H; 3 C120Y/−, 4 WT littermates in (I)). J Representative photographs of thymi from 10- or 20-wk-old Foxn1-Cre;Ovol2^fl/fl mice and their Ovol2^fl/fl or Foxn1-Cre;Ovol2^+/+ littermates, and age matched Ovol2^C120Y/− mice. (K) Representative hematoxylin and eosin (H&E) (left) and Oil red O (right) staining of Ovol2^C120Y/−, Foxn1-Cre;Ovol2^fl/fl, or age matched WT thymi at 20 wk of age. Black arrows indicate adipose tissue. L, M Numbers (L) and frequencies (M) of thymocytes in Foxn1-Cre,Ovol2^fl/fl and Ovol2^fl/fl littermates at 20 weeks of age (n = 4 Foxn1-Cre,Ovol2^fl/fl, 7 Ovol2^fl/fl littermates). N Surface marker CD44 expression by thymocytes in 20-wk-old Foxn1-Cre;Ovol2^fl/fl and Ovol2^fl/fl littermates (n = 4 Foxn1-Cre,Ovol2^fl/fl, 7 Ovol2^fl/fl littermates). O Quantitative analysis of the ova-specific cytotoxic T cell killing response in 20-wk-old Foxn1-Cre;Ovol2^fl/fl mice and WT littermates immunized with alum-ova (n = 5 Foxn1-Cre,Ovol2^fl/fl, 17 Ovol2^fl/fl littermates). P Representative flow cytometry plots of TECs from 20-wk-old Foxn1-Cre;Ovol2^fl/fl mice and their Ovol2^fl/fl littermates, and age matched Ovol2^C120Y/C120Y mice. Numbers adjacent to outlined regions indicate percent cells in each (mean ± SD) (n = 5 C120Y/C120Y mice, 4 Foxn1-Cre,Ovol2^fl/fl mice, 6 age matched WT mice). Data are representative of two independent experiments (F, G, I, L–O). Data points represent individual mice (F, G, I, L–O) and means ± SD are indicated. P-values were determined by two-tailed Student’s t-test (F, G, I, L–O). MFI Mean fluorescence intensity. +/+, WT; C120Y/−, Ovol2^C120Y/−; C120Y/C102Y, Ovol2^C102Y/C120Y; and fl/fl, Ovol2^fl/fl.