Fig. 4.

Mint deficiency enhances ETP production and impairs the DN1–DN2 transition in in vitro T cell differentiation. (A) The cells cultured on Tst-4/Dll1 (+ Dll1) or Tst-4 (No Dll1) were analyzed for their expression of Thy1.2 and CD19 at various time points. The cells generated on Tst-4/Dll1 were analyzed for their expression of lineage markers or CD4 and CD8 at various time points, and the absolute cell numbers of Lineage-negative (DN) cells and DP cells are calculated. (B) The cells generated in A were analyzed for their expression of TCRβ and TCRγδ at day 14, and the absolute cell numbers of αβT and γδT cells are calculated. (C) The Lin⁻ cells among the cells generated in A were analyzed for their expression of CD44 and CD25 at indicated day in vitro, and they were subdivided into DN1–DN4 subsets as in Fig. 2B. (Upper) Representative FACS profiles and (Lower) absolute cell numbers of indicated thymocyte subsets. (D) The Lin⁻ cells among the cells generated in A were analyzed for their expression of CD44, CD25, and c-Kit at day 6, and the absolute cell numbers of ETPs are calculated. Each value represents the mean ± SEM of three independent samples per group. ∗, P < 0.05 by Student's unpaired t test.