STIM2-mediated SOCE regulates activation of NF-κB. (A-B) Analysis of calcium-dependent activation of NF-κB and upstream signaling pathways in WT neutrophils. Neutrophils were stimulated with zymosan (100 μg/mL) in the presence or absence of BAPTA-AM to chelate available calcium. Nuclear and cytoplasmic fractions were immunoblotted for the indicated proteins. (C) Analysis of NF-κB activation and upstream signaling pathways in WT, Stim1^VavCre, Stim2^VavCre, and Stim1/2^VavCre neutrophils stimulated with zymosan (100 μg/mL). Nuclear and cytoplasmic fractions were blotted for the indicated proteins. n.s., nonspecific band. Blots are representative of more than 3 independent experiments. (Right) Mean band intensities (normalized to WT) ± SEM compiled from 3 to 4 independent experiments. (D) Analysis of phospho-CAMKII in cytoplasmic fractions of WT, Stim1^VavCre, Stim2^VavCre, and Stim1/2^VavCre neutrophils stimulated with zymosan (100 μg/mL). (Right) Mean band intensities (normalized to WT) ± SEM compiled from 3 to 4 independent experiments. Cyto., cytoplasmic.