Figure 4

Calcium requirement of TLR4-stimulated β-END release. (A) Release of β-END in the supernatant was quantified after incubation of CD14⁺ monocytes for 15 min with 10 μM ionomycin (iono) or medium (control) (n = 3, * p < 0.05, paired t-test). (B) CD14⁺ monocytes were stimulated as described before and stained on cytospins using a ββ-END antibody (red) and confocal analysis (representative example, n = 3). (C-E) CD14⁺ monocytes were preincubated with BAPTA-AM (100 μM, n = 24–28, * p < 0.05, One Way RM ANOVA, Student-Newman-Keuls, C), 2-APB (100 nM, n = 11 * p < 0.05, One Way RM ANOVA, Student-Newman-Keuls, D) for 10 min before the addition of 10 μg/ml LPS for 15 min. Secretion of β-END was quantified in the supernatant. All data are presented as MEAN ± SEM.