Additional file 1: Figure S1. of Differential Lyn-dependence of the SHIP1-deficient mast cell phenotype

Impaired Ca2+ mobilization in Lyn-/- BMMCs is independent of the stimulus concentration. Wt and Lyn-/- BMMCs were preloaded and starved overnight with 0.15 μg/ml IgE. Ca2+ mobilization was measured for 4 min by flow cytometry using the Ca2+-sensitive fluorescent dyes fluo-3 (Ca2+ b) and fura red (Ca2+ ub). Steady-state fluorescence was assessed for 1 min before BMMCs were stimulated with the indicated Ag concentrations. The arrow marks the time point of stimulus addition. Comparable results were obtained with cells from different BMMC cultures (n = 3). (PDF 289 kb

Additional file 4: Figure S4. of Differential Lyn-dependence of the SHIP1-deficient mast cell phenotype

Comparison of proinflammatory cytokine mRNA production in optimally Ag-stimulated wt, Lyn-/-, SHIP1-/-, and dko BMMCs. Wt, Lyn-/-, SHIP1-/-, and dko BMMCs were preloaded and starved overnight with 0.15 μg/ml IgE. BMMCs were left unstimulated (con) or stimulated with 20 ng/ml of Ag for 90 min. The amounts of Il6 mRNA (A) and Tnf mRNA (B) were determined by RT-qPCR using the Pfaffl method. Data obtained by analyzing cells from independent cultures are shown (the results already shown in Figs. 2c & d are indicated). (PDF 296 kb