Fig. 2
From: AKT2 deficiency impairs formation of the BCR signalosome
AKT2 deficiency impairs B cell spreading, BCR cluster formation and BCR signalosome during B cell early activation. Spleen cells from 8-week old WT and AKT2 KO mice (n = 3) were incubated with biotinylated Fab fragment goat anti-mouse IgM and IgG, labeled with Alexa Fluor 546 (AF546-mB-Fab’-anti-Ig) tethered to lipid bilayers. After incubation at 37 °C for varying lengths of time, cells were fixed, permeabilized, and stained with anti-pY and anti-pBtk antibodies. Shown are the respective images by TIRFm (a, b), and the B-cell contact zone area (c), mean fluorescence intensity (MFI) of BCR (d), pY (e) and pBtk (f) in the contact zone area were analyzed using IRM and TIRFm. Data were generated from > 50 cells from 3 individual experiments, and presented as mean values ± SEM, Scale bars, 2.5 μm. *p < 0.05