Fig. 7.

The effect of EGTA treatment on the spatiotemporal characteristics of GFP-PKC-βII in HEK 293 cells after KISS1R activation. A–F, Representative images selected from a time series of laser-scanning confocal microscopic images showing the plasma membrane and cytosolic localization of GFP-PKC-βII in KISS1R-expressing HEK 293 cells after 100 nm Kp-10 treatment. Approximately 4 min Kp-activated cells were treated with 2.5 mm EGTA. White solid arrow shows translocation of GFP-PKC-βII (panel B). Prolonged exposure to EGTA caused cells to lift due to the chelation of extracellular Ca²⁺; as a result, cells could only be imaged for up to 10–25 min after EGTA treatment. These studies were conducted five independent times, and representative images from one independent experiment are shown. ms, Millisecond; s, second.