Fig. 1: Equivalent immunodeficiencies in mice lacking shieldin and CST.
a, Schematic representation of the Shld2 and Shld3 (knockout) and Ctc1 (conditional) alleles generated in this study. TSS, transcription start site. b, Absolute numbers of B220+ B cells in the bone marrow (one femur and one tibia) and spleen (n = 4–8 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.). NS, not significant. c, Absolute numbers of B cell precursors (Hardy fraction A, B220+CD43+BP1−CD24−; Hardy fraction B, B220+CD43+BP1−CD24+; Hardy fraction C, B220+CD43+BP1+CD24+; Hardy fraction D, B220+CD43−IgM−IgD−; Hardy fraction E, B220+CD43−IgM+IgD−; Hardy fraction F, B220+CD43−IgM+IgD+) in the bone marrow (one femur and one tibia) (n = 4–8 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.). d, 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 analysis of variance (ANOVA) with Tukey’s correction (mean ± s.e.m.). e, CTV-labeled splenic B cells were stimulated as indicated and stained for surface IgG1 after 96 h. Representative data, n > 6 mice. f, 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.). g, CTV dilution in purified B cells cultured in the presence of LPS and IL-4 for 96 h. Representative data, n > 6 mice. h, NP-specific serum IgM (left) and IgG1 (right) at indicated times after NP-CGG immunization. Representative data, n = 2 independent experiments, each with four mice. Significance was determined by an unpaired two tailed t-test (mean ± s.e.m.). Gating strategies for the above flow cytometry data are provided in Supplementary Fig. 1.
