Figure 6.

Silencing STIM1 expression inhibits Ca²⁺ mobilization, degranulation and cytokine production in LAD2 mast cells. LAD2 cells were stably transduced with lentivirus containing STIM1 or scrambled shRNA (control). (A) A representative blot of STIM1 levels in wild type (WT), scrambled and STIM1 shRNA-transduced cells is shown. (B) Control and STIM1 knockdown (KD) cells were labeled with the Calcium 6 dye in Ca²⁺-free buffer. Traces represent the changes in fluorescence following the addition of CST-14 (300 nM) and 2 mM Ca²⁺. (C,D) Cells were stimulated with indicated concentrations of CST-14 or LL-37 and β-hexosaminidase release was determined. (E,F) Cells were treated with CST-14 for 6 h, and ELISA was performed to estimate TNF-α and IL-2 levels in the supernatants. Data is mean ± S.E. from three experiments. Statistical significance was determined by two-way ANOVA. *p < 0.05 and **p < 0.01.