Extended Data Fig. 4: Functional PI3Kγ is required for optimal ASC differentiation in vitro and in vivo despite being dispensable for innate-like B cell IgM responses.
From: PI3Kγ in B cells promotes antibody responses and generation of antibody-secreting cells
(a) Expression of PI3K genes in human/mouse immune cells derived from Immgen3. (b) Expression of Pik3cg in mouse B cells comparing splenic follicular and germinal center B cells to innate-like marginal zone and B1a B cells. (c-d) Mice were immunized i.p. with TI antigens and serum was taken on day 5 to measure antigen-specific IgM via ELISA. (e-f) Antigen-specific ASCs were measured from splenocytes via ELISPOT in WT mice treated as described for Fig. 4e, f. (g-h) Assessment of in-vitro differentiation of naïve mouse B cells into ASCs (anti-CD40, IL-4, IL-5 gating on CD138+ after 4 days) in utilizing B cells from Blimp1-YFP mice treated in vitro with DMSO vehicle or the PI3Kγ inhibitor IPI-549. Data are from 2-4 independent experiments. Data presented as box-and-whisker plots show median, min, and max. (c) n = 11 for controls, n = 12 for KOs. (d) n = 12 for both groups. (e) n = 15 for both groups. (f) n = 13 for both. (g-h) n = 6 for each group. Each dot represents different biological samples and statistical analysis was performed using non-parametric two-tailed unpaired T-tests, with the exception of panel H which used non-parametric paired T-test.
