Figure 6. NHERF1 and NHERF2 contribute to C3a-induced degranulation response in LAD2 and CD34⁺ primary human mast cells.

LAD2 or CD34⁺ primary human mast cells were stably transduced with scrambled shRNA control lentivirus or shRNA lentivirus targeted against NHERF1 or NHERF2. Western blotting was performed to assess NHERF expression levels in shRNA control and NHERF KD (A) LAD2 and (C) CD34⁺ primary human mast cells. A representative blot from three independent experiments is shown. shRNA control or NHERF KD (B) LAD2 mast cells or (D) CD34⁺ mast cells were stimulated with indicated concentrations of C3a or CST (100 nM) and percent degranulation (β-hexosaminidase release) was determined. LAD2 cells were also stimulated by NP-specific IgE/NP-BSA (Ag) as a non-GPCR control. Data are mean ± SEM of three experiments. Statistical significance was determined by two-way ANOVA with Bonferroni's post test. ** indicates p<0.001.