Figure 4.

Subcellular localization of the EGFP tagged NEDD4L-C2(+) isoform and the NEDD4L C2 domain (C2(only)) in response to ionomycin and Ca²⁺ treatment. Confocal images of live A6 cells transiently transfected with EGFP-NEDD4L-C2(+) (A-C) or EGFP-C2(only) (D-F) that were incubated in 1.97 Ca²⁺. Images were acquired at 0 (A), 40 (B), 90 (C), 0 (D), 10 (E), and 30 (F) seconds after a 10 uM ionomycin addition. Both EGFP-NEDD4L-C2(+) and EGFP-C2(only) respond to the Ca²⁺ stimulus by first mobilizing to the plasma membrane and then relocalizing to numerous small intracellular structures. Scale bars are equivalent to 10 μm.