Figure 2: Setd1-interacting domain of Cxxc1 is required for intrathymic T-cell development.

(a) DN3 cells from Cxxc1-deficient mice were sorted and transfected with indicated retrovirus. GFP⁺ thymocytes were analysed by fluorescence-activated cell sorting after co-culturing with OP9-DL1 for 5 days. Data comes from four independently sorted DN3 samples (each from three mice). (b) Detection of total H3K4me3. Crude histone protein were extracted from DP thymocytes sorted from control or Cxxc1-deficient mice using acid extraction, the protein was subjected to H3K4me3 ELISA analysis. Data comes from three separately sorted DP cell sets (each from two mice). The statistical significance was calculated by unpaired t-test (two tailed). **P<0.01. Error bars indicate s.d. (c) Detection of Setd1 protein level. DP thymocytes from Cxxc1-deficient and control mice were used for western blot analysis. Number under lanes indicate densitometry of Setd1 (relative to Actb at same lane, below). Data comes from three separately sorted DP cell sets (each from three mice). (d) Dot blotting showed total 5 mC levels in sequentially diluted genomic DNA from indicated DP thymocytes. Data are representative of three separate experiments (each from two mice).