Figure 3.

Flagellin-induced ATP release and calcium mobilization are Toll dependent. NCIH292 cells transfected with empty vector (EV) or MyD88 DNM were exposed to α-ASGM1 antibody (1:100) or flagellin (Flgn, 10 μg/ml) for times indicated. (A) ATP release into the extracellular medium was monitored using a luciferin/luciferase assay. ATP release was inhibited by DNM MyD88. (B) Ratiometric Ca²⁺ imaging of islands of NCIH292 cells loaded with Fura-2/AM show increases in the 340/380-nm excitation fluorescence ratio (510 nm emission) ∼ 20 s after exposure to flagellin (B, bottom left panel). This increase in Ca²⁺ mobilization is reduced in the presence of DNM MyD88 (B, bottom right panel). Quantification of results from three separate experiments is shown in the histogram.