Extended Data Fig. 1: Generation and characterization of Runx3WRPW mouse strain.
(a) Image of an agarose electrophoresis gel of Bcl1-digested PCR products of nine one-week-old F0 founders on a B6 background generated by CRISPR/Cas9 genome editing to generate the Rx3W allele. Predicted nucleotide sequences for WT(WRPY) and mutant WRPW alleles and the Bcl1 enzyme site are shown at the bottom. The DNA sequence of the PCR product of an F0 founder on an ICR background, through which Rx3W strain was established, is shown at the right. PCR products were cloned into the plasmid and sequenced by the Sanger method. A single experiment was performed. (b) Pseudo-color plots showing CD3ε and NK1.1 expression by spleen cells. The graph at the bottom shows a statistical summary of the frequency of NK1.1+ cells in CD45+ spleen cells. Rx3+/+ (n = 5), Rx3+/W (n = 5) and Rx3W/W (n = 5). Mean ± SD. *: p < 0.05, ***: p < 0.005, ****: p < 0.0001 (one-way ANOVA with Tukey’s multiple comparison). (c) Pseudo-color plots showing CD45 expression by epidermal cells, MHC-II and CD3ε expression by the CD45+ subset, and EpCAM and CD24 expression in the CD45+ MHC-II+ subset. The graph at the bottom shows a statistical summary of the frequency of the indicated cells in the indicated population. Rx3+/+ (n = 3), Rx3+/W (n = 3) and Rx3W/W (n = 3). Mean ± SD. *: p < 0.05, **: p < 0.01, ***: p < 0.005, ****: p < 0.0001 (one-way ANOVA with Tukey’s multiple comparison).
