Tyrosine 223 to phenylalanine substitution in BTK does not affect B-cell development in bone marrow or peripheral B-cell subsets in spleen and peritoneal cavity. (A-C) (left) Gating strategy for identification of indicated B-cell subsets. Prior gating was performed as indicated above the fluorescence-activated cell sorting plots. Propidium iodide (PI) was used for dead cell discrimination. (A-B) (right) Absolute numbers of total B cells and B-cell subsets in bone marrow (BM) and spleen. (C) (right) Frequency of subsets within B cells in the peritoneal cavity (PeC). In (A-C) (right), each dot represents data from individual animals, vertical bars indicate mean and 2-tailed Mann-Whitney test was used to calculate significance. Immature B cell (immB); mature B cell (matB); transitional 1 B cell (T1); transitional 2 B cell (T2); marginal zone B cell (MZ); follicular B cell (FoB); switched B cell (Sw). None of the comparisons yielded any significant difference.