Supplementary Figure 1: Basal characterization of Pik3cdE1020K/+ mouse model. | Nature Immunology

Supplementary Figure 1: Basal characterization of Pik3cdE1020K/+ mouse model.

From: Hyperactivated PI3Kδ promotes self and commensal reactivity at the expense of optimal humoral immunity

Supplementary Figure 1

a, Schematic representation of Pik3cdE1020K/+ generation. After crossing to ZP3-Cre mice to remove the neoR cassette, the point mutation and one LoxP site in the upstream intron remain. (b, d-h) Wild-type and Pik3cdE1020K/+ mice were analyzed at steady state between 2 and 4 months of age. b, Cellularity of resting lymph node (n=5 per group). c, Representative contour plots of CD44 and CD62L on CD4+ T cells in the spleen of 1-year-old mice. d, Numbers of CD93+CD138 transitional B cells in the spleen (percent of B220+CD19+); representative flow staining and histograms of percentages and numbers of T1, T2, T3 and IgM CD23 B cells (gated on B220+CD19+CD93+CD138) (wild-type n=3, Pik3cdE1020K/+ n=4). e, Representative FACS plots of surface IgM and IgD (gated on B220+CD19+ B cells in the spleen) and relative histograms (wild-type n=8 Pik3cdE1020K/+ n=7). f, Illustrative contour plots for CD21 and CD23 to identify MZ, FO, and CD21CD23 B cells in the spleen (percent of B220+CD19+). Bar graph on the right (wild-type n=6, Pik3cdE1020K/+ n=5). (g-h) Analysis of peritoneal cavity (n=3 per group). g, Cellularity of peritoneal lavage; representative flow staining and cell counts of CD3+ T and CD19+ B cells. h, Representative contour plots and cell counts of B1 (CD23) and B2 (CD23+) B cells gated on CD19+ cells as in (g); on the right, representative histograms and numbers of B1a (CD5+) and B1b (CD5) gated on B1 B cells. Data in (b-f) are representative of three, and data in (g,h) of two, independent experiments. Data are represented as mean ± SEM with each dot indicating one mouse. Significance analyzed by Mann-Whitney U test. *P < 0.05; **P < 0.01; ***P < 0.001