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. 2016 Oct 23;2016:2048987. doi: 10.1155/2016/2048987

Figure 4.

Figure 4

Maturation of mast cells by GSK-3β inhibitor. (a)–(e) BMMCs were cultured for 20 days under two conditions: one with IL-3 (1 ng/mL), SCF (100 ng/mL), and BIO and the other with only IL-3 (1 ng/mL) and SCF (100 ng/mL) (control). (a) Suspensions of mast cells were stained with fluorescence-labeled anti-FcεRI, anti-c-kit, and anti-CD81 antibodies for 30 min on ice. 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) Granule protease activities of mast cells were measured. (e) β-Hexosaminidase release from cells after stimulation with compound 48/80. All data represent the means ± SD (n = 4). ∗ p < 0.05.