Fig. 3

Lyn deficiency is dominant to combined SHIP1 deficiency concerning degranulation and relevant signaling events. Wt, Lyn-/-, SHIP1-/-, and dko BMMCs were preloaded and starved overnight with 0.15 μg/ml IgE. a Unstarved BMMCs were left untreated (con) or stimulated with Ag (20 ng/ml) for 20 min. Degranulation was determined by β-hexosaminidase assay. Each point is the mean of triplicates ± SD. For the response to different Ag concentrations see Additional file 5: Figure S5A. b Ca²⁺ mobilization was measured in wt, Lyn-/-, SHIP1-/-, and dko BMMCs for 6 min by flow cytometry as described in Fig. 1b. For the response to different Ag concentrations see Additional file 5: Figure S5B. c BMMCs were left untreated (-) or stimulated with Ag (20 ng/ml) for the indicated time points. Whole-cell lysates were subjected to WB analysis with anti-P-Tyr (upper panel) and anti-GAPDH (loading control, lower panel) antibodies. d BMMCs were left untreated (-) or stimulated with Ag (200 ng/ml) for the indicated time points. Whole-cell lysates were subjected to WB analysis with antibodies against P-PLC-γ1 (top panel), P-LAT1 (middle panel), and p85 (loading control, bottom panel). e BMMCs were treated as in (d). Whole-cell lysates were subjected to WB analysis with antibodies against P-ERK1/2 (top panel), P-p38 (middle panel), and p85 (loading control, bottom panel). Densitometry was performed and relative expression levels are indicated under each band. Comparable results were obtained with cells from different BMMC cultures (n = 3 (b, d); n = 2 (c); n > 3 (a, e))