Cholesterol enrichment attenuates the ATP agonist calcium response. Prior to the experiment, cells were incubated with PBS with Ca+2 for 30 min (solid line), 3.5 mM water-soluble cholesterol for 30 min (dashed line) or 50 μM of SOC inhibitor SKF-96365 for 10 min (dotted line). (a) Ca+2 response of each condition to 500 µL of 100 µM ATP dissolved in PBS with Ca+2. (b) Bar graph representing average responses for each condition at the peak response, 60 and 100 s. Cholesterol treated cells show a reduced transient and sustained calcium response, which was statically significant from unenriched and SKF treated cells. Average responses represent multiple coverslips (cs) and total cell count among coverslips (n) Unenriched: #cs = 8, n = 512; SKF: #cs = 5, n = 298; Cholesterol: #cs = 7, n = 441. Mean and SEM were plotted two-tailed t test p < 0.0001.