FIGURE 7:

SHIP1 recruitment is absent in RBL-Lyn^KO cells. (A) RBL-WT or (B) Lyn^KO cells were transfected to express SHIP1-mNG (green) and incubated overnight with IgE-AF647 (magenta). Cells were activated with the addition of 0.1 µg/ml DNP₂₅-BSA and confocal images collected at specified time points. SHIP1-mNG recruitment is not observed in the absence of Lyn. (C, D) RBL-WT cells coexpressing SHIP1-mNG (green) and either (C) TT-β or (D) TT-γ. Aggregation of the anti-Tac primary antibody (magenta), with 25 µg/ml anti-mouse secondary antibody, induced SHIP1-mNG recruitment to TT-β but not TT-γ. Representative image shown of 103 (A), 107 (B), 59 (C), and 38 (D) cells imaged. See Supplemental Figure S1 for quantification of SHIP1-mNG recruitment frequency. Scale bar is 5 µm.