Fig. 3: Pol ζ cooperation with shieldin supports NHEJ in CSR.
a, Schematic representation of the Rev3l conditional allele. b, Absolute numbers of B220+ B cells in the bone marrow (one femur and one tibia) and spleen (n = 4–6 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two tailed t-test (mean ± s.e.m.). c, CTV dilution in purified B cells cultured in the presence of LPS and IL-4 for 96 h. Representative data, n > 6 mice. d, IgG1+ B cells as a proportion of total B cells (%) for each cell generation as determined by CTV staining and proliferation-associated dye dilution (n = 4–6 mice per genotype, where each data point is a single mouse; mean ± s.e.m.). e, Splenic B cells cultured with the indicated stimuli (96 h) and stained for surface IgG1, IgE, IgG2b or IgG3 (n = 4–7 mice per genotype, where each data point is a single mouse). CSR 100%, mean immunoglobulin isotype switch frequency of two control animals in each experiment. Significance was determined by a two-way ANOVA with Tukey’s correction (mean ± s.e.m.). f, CTV-labeled splenic B cells were stimulated as indicated and stained for surface IgG1 after 96 h. Representative data, n > 6 mice. g, Splenic B cells cultured with the indicated stimuli (96 h) and stained for surface IgG1, IgE, IgG2b or IgG3 (n = 4–7 mice per genotype, where each data point is a single mouse). CSR 100%, mean immunoglobulin isotype switch frequency of two control animals in each experiment. Significance was determined by a two-way ANOVA with Tukey’s correction (mean ± s.e.m.). h, Splenic B cells cultured with the indicated stimuli (96 h) and stained for surface IgG1, IgE, IgG2b or IgG3 (n = 4–7 mice per genotype, where each data point is a single mouse). CSR 100%, mean immunoglobulin isotype switch frequency of two control animals in each experiment. Significance was determined by a two-way ANOVA with Tukey’s correction (mean ± s.e.m.).
