Figure 3
Divalent cation transport activities in multi-well plates. A- C. Apoaequorin bioluminescence for cytosolic Ca2+ , Mg2+ and Mn2+, respectively. Cell cultures were distributes in 0.1 ml volumes in 96-well plates at 0.1 OD600 densities. A bolus of 0.1 ml 1 M of CaCl2, MgCl2, or MnCl2 was added at 5 seconds and the relative light units (RLUs) were recorded every 0.5 second for 60 seconds. The signals for experimental group were normalized against the RLU of the 5 second point for that group (RLUt (0 .. 22)/RLUt = 5) and the signals for the first 22 seconds are presented. D. Dual magnesium and calcium transport. Cell cultures in 0.08 ml volumes were distributed in 96-well plates at 0.1 OD600 density. MgCl2 was added (1M, 0.02 ml) at 2 seconds followed by 0.1 ml of 1M CaCl2 at 5 seconds. RLUs were normalized as in A–C. Data presented is representative of four experiments for A–C and three experiments for D.