Calcium mobilization, cytokine production and PGE₂ secretion are induced by mannitol in LAD2 cells.

<p>Determination of calcium flux following mannitol stimulation (10%) was performed in LAD2 cells loaded with Fluo-4 dye, either with calcium containing media (A) or without extracellular calcium (B), as described in material and methods section. After defining basal conditions the stimuli was added (time 0) and measured 10 minutes. Real time polymerase chain reaction was performed in LAD2 cells that were either unstimulated or stimulated with 10% mannitol, using interleukin-8 and tumor necrosis factor alfa as probes. PMA+ Ionomycin (Sigma) were used as positive control stimuli (C). PGE₂ release was measured at different times as indicated in the figure using an ELISA assay. Results are expressed in pg per 1×10⁶ cells (D). Results are triplicates expressed as mean ± standard deviation, and are representative of three independent experiments. Statistical significance (* p≤0.05, **p≤0.01) is relative to unstimulated control cells. RFU: relative fluorescence units.