Figure 4. NaCl plus FK506 treatment caused a slow and persistent increase in the cytoplasmic Ca²⁺ level.

The wild-type (A), Δppb1 (B) or Δprz1 (C) cells harboring pKB6892 were cultured as described in Figure 2B. A 10 µl volume of distilled water or 2 M stock of NaCl were added into the 96-well plate, and then 90 µl volume of cells were rapidly added to each sample. The cells were incubated for 30 min to reach a steady-state cytoplasmic Ca²⁺ level and the luminescence was followed for 60 min. Then 2 µl of distilled water or FK506 with a concentration of 50 µg/ml were added to the samples and the luminescence was monitored up to 240 min. Left panel: The data shown are representative of multiple experiments. Right panel: The data represent the means ± standard deviations of RLU taken at 240 min from three independent experiments, and each sample was analyzed in duplicate.