Wnt-β-Catenin Signaling Promotes the Maturation of Mast Cells

Figure 2

Maturation of mast cells by Wnt5a. (a–e) BMMCs were cultured for 20 days under two conditions: one with IL-3 (1 ng/mL), SCF (100 ng/mL), and Wnt5a (50 ng/mL) and the other with only IL-3 (1 ng/mL) and SCF (100 ng/mL) (control). (a) BMMCs were stained with fluorescence-labeled anti-FcεRI, anti-c-kit, and anti-CD81 antibodies for 30 min on ice. The stained cells were washed, resuspended in 1% FBS-PBS, and analyzed by flow cytometry. Surface marker expression was analyzed by gating on viable cells in the FSC/SSC. One representative dot plot out of five independent experiments is shown. (b) Gene expression levels of HDC, MCP5, and CPA were measured by quantitative RT-PCR. Gene expression levels in the control were taken as 1.0. (c) β-Hexosaminidase content was also measured. (d) Cell extracts prepared from BMMCs were assayed for tryptase and CPA activities as described in Section 2. (e) β-Hexosaminidase release from cells after stimulation with compound 48/80. All data represent the means ± SD (). .

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