Figure 4.

HgCl2 regulates Ca²⁺ mobilization leading to secretion of IL-1β and IL-18. (A) [Ca²⁺ ]i levels in treated with/without HgCl2. BMDMs from C57BL/6 were loaded with calcium indicator dye Fluo-4AM and treated with/without HgCl2 (20μM). Live [Ca²⁺ ]i levels were continuously measured using confocal spinning disc microscopy for 40 minutes. Ionomycin (1μM) was added at 30min. The average Mean Fluorescence Intensity (MFI) (Y-axis) from 5 independent fields was measured and plotted against time on the X-axis. The data shown is a representative graph of 2 experiments. BMDMs from C57BL/6 mice were treated with LCWE (10μg/ml) and treated with/without HgCl2 (20μM). Prior to the treatment of HgCl2, cells were treated with/without Ca²⁺ inhibitors XeC (5μM), Tg (100nM), U73122 (10μM), and 2-APB (100μM). (B) Levels of IL-1β from supernatants of BMDMs, measured by ELISA (n=3), *p<0.05. (C) Levels of IL-18 from supernatants, measured by ELISA (n=3), *p<0.05. Statistical analysis between cultures of LCWE + HgCl2 - stimulated BMDM with/without Ca²⁺ inhibitors is done using the Unpaired t-test.