Figure 2.
The recruitment of pY, pBtk, and pSHIP to BCR clusters in B cells stimulated by sAg is reduced in Mst1 KO B cells. Splenic B cells were incubated with AF546–mB-Fab′–anti-Ig without (−) or with sAg at 4°C, washed, and warmed to 37°C for varying lengths of time. After fixation and permeabilization, the cells were stained for pY, pBtk, and pSHIP and analyzed using CFm (A,D,G). The MIF of pY, pBtk, and pSHIP was generated using NIS-Elements AR 3.2 software (B,E,H). The Pearson’s correlation coefficients between BCR and pY, pBtk, or pSHIP staining in sAg-stimulated cells were determined using NIS-Elements AR 3.2 software (C,F,I). Shown are representative images at indicated times and the average values (±SD) of ∼50 cells from 3 independent experiments. Scale bars, 2.5 μm. *P < .01.