Fig. 7. ELMO is involved in CaSR-induced PM ruffling. (A) Expression of GFP-ELMO(T629) but not GFP-WT-ELMO impairs CaSR-induced PM ruffling. CaSR-HEK cells transfected with GFP-ELMO(T629), GFP-WT-ELMO or GFP were stimulated with 5 mM CaCl₂ for 10 minutes, fixed and stained for GFP (green) and F-actin (red). The micrographs are representative of three independent experiments. Arrowheads indicate cells exhibiting ruffles and arrows indicate cells not ruffling. Right panels are a magnified view of the boxed regions in the merge panels. (B) Quantification of cells presented in (A). ^*P0.001. NS, not significantly different (P>0.05) (n=3). (C) GFP-ELMO and Flag-CaSR colocalise in protrusions. HEK cells co-expressing the two plasmids indicated above were challenged with 5 mM CaCl₂ for 2 or 10 minutes, fixed and stained for GFP-ELMO (green) and Flag-CaSR (red). Arrowheads show protrusions, and colocalisation areas appear in yellow. Bar, 5 µm.