Fig. 5

The phosphorylation of CD19 is decreased during B cell early activation in AKT2 KO mice. The phosphorylation of CD19 and Btk during B cell activation were determined by CFm and flow cytometry. a Spleen cells from WT and AKT2 KO mice (n = 3) were seed on the lysine coated slides, and incubated with AF546-mB-F (ab)₂′-anti-Ig and streptavidin on ice. Cells were stimulated at 37 °C for indicated times, and then fixed and permeabilized, following stained with monoclonal antibodies of pCD19 and pBtk. Shown are the representative images of CFm (a, b). The Pearson’s correlation coefficients of pBtk and pCD19 at different time points during activation were analyzed by NIS-Elements AR 3.2 software (c). Data were generated from more than 50 cells from 3 independent experiments. Scale bars, 2.5 μm. Flow cytometry analysis of the MFI of pCD19 and pBtk of B cells stimulated by sAg (d, e). CD19 expression of FO B cells (B220⁺ IgD⁺ IgM^low) and MZ B cells (B220⁺ CD21⁺ CD23^low) was detected by flow cytometry (f). 3 independent experiments were performed, and data were presented as mean ± SEM, *p < 0.05