Supplementary Figure 7: Overlapping functions of LRF and ThPOK in CD4 expression.

(a) Histogram overlays show CD4 expression on sorted CD4^lo (1), CD4⁺ CD8^– (2) CD44^lo spleen T cells from Thpok^pd mice or their Thpok^+/+ CD4⁺ CD8^- counterparts (3) ex vivo (left panel), or after 5 d culture under T_H1 conditions (right). Representative of three experiments. (b,c) Contours plots assess 6–8-week-old littermates of the indicated genotype for expression of CD4 and CD8 on total (left) and TCRβ^hiCD24^lo (right) thymocytes (b), or (c) on total (left) or gated TCR⁺ YFP⁺ CD44^lo (right) spleen T cells. Figures are representative of three experiments, summarized in d. (d) Plot summarizes numbers (× 10⁶) of mature (TCR^hi CD24^lo) CD4 and CD8SP thymocytes (top) and of CD4⁺ (including CD4^lo and pDP) and CD8⁺ T cells from 6–8 week-old mice of the indicated genotype. Significance was determined by Student’s t-test (*P < 0.05, ** P < 0.01; not significant otherwise). (e) Immunoblot analysis of Runx protein expression in effector T cells obtained after 5-d culture under TH1 conditions from naïve CD8⁺ T cells from Thpok^+/+ Zbtb7a^+/+ mice or CD4⁺ T cells from Thpok^pd Zbtb7a^+/+ or Thpok^pd Zbtb7a^pd mice. The same membrane was subsequently stripped and reprobed for β-actin expression to evaluate lane loading. Representative of two immunoblots.