Fig. 2. Characterization of Dapl1 reporter.

(A) CRISPR-Cas9–mediated Dapl1^ZsG reporter gene construct strategy. 5′UTR, 5′ untranslated region. (B) Dapl1 expression in peripheral lymph nodes of Dapl1^ZsG/WT mice (n = 3; left), with proportions of Dapl1⁺ cells and Dapl1⁺ MFIs of CD4 T cells (light green), CD8 T cells (green), or CD4⁻CD8⁻ cells (DN, gray) (right). (C) WB of Dapl1 and Zap70 expression in cell lysates from CD8 T cells from lymph nodes of WT, Dapl1^ZsG/WT, or Dapl1^ZsG/ZsG mice. (D) Representative UMAP projection (left) of multicolor flow cytometry data from spleens of Dapl1^ZsG/WT mice with ZsG fluorescence. DC, dendritic cell; pDCs, plasmacytoid dendritic cells. (E) ZsG and RFP expression on splenocytes from a Dapl1^ZsG/WTFoxp3^RFP/RFP mouse. (F to H) Proportion of ZsG expressing cells within (F) spleen, (G) liver, and (H) gut subsets: small intestine intraepithelial lymphocyte (IEL), small intestine lamina propria lymphocyte (LPL), colon IEL, or colon LPL. Statistical analyses were calculated using ordinary one-way ANOVAs with Tukey’s correction for multiple comparisons. Data are representative of more than three independent experiments [(B), (E), and (F)], one experiment [(C) and (D)], three biological replicates from one experiment (G), or at least two biological replicates pooled from three independent experiments (H).