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. 2019 Jul 16;216(9):2170–2183. doi: 10.1084/jem.20181953

Figure 1.

Figure 1.

Itch limits serum antibody and activated B cells. (A) Quantity of total antibody in serum of WT and Itch KO mice was determined by ELISA (mice between 8 and 12 wk old, n = 4–8, compiled from two independent experiments, multiple t tests, Holm–Sidak correction). (B) Anti-dsDNA IgG in serum was determined by ELISA (mice between 12 and 24 wk, n = 9–13, compiled from three independent experiments, two-way ANOVA). Serum from New Zealand black × New Zealand white F1 mice was included as a positive control (n = 2). (C and D) Preimmune and activated B cell subsets in spleen were quantified by flow cytometry, and representative flow cytometry plots are shown. Cells were gated on live singlets, and then transitional B cells were CD19+CD93+, FO B cells were CD19+CD93−CD23+, and marginal zone B cells were CD19+CD23lo/−CD21+. For GC and class-switched non-GC B cells, cells were first gated on IgD−CD4−CD8−F480−GR1−CD19+IgM−, and then GC B cells were GL7+CD38−, and class-switched non-GC B cells were GL7− (n = 6–8, compiled from two independent experiments, Mann–Whitney test, WT mice are C57Bl/6). (E) Total IgM-, IgG1-, and IgG2c-secreting PCs were enumerated by ELISPOT (n = 8, Mann–Whitney test, compiled from two independent experiments; error bars indicate SEM; WT mice are C57Bl/6). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.