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Figure 6


Fig. 6. ARF6 is involved in CaSR-induced PM ruffling. (A) ARF6(T27N) impairs CaSR-induced PM ruffling. CaSR-HEK cells transfected with GFP, GFP-ARF6T27N or GFP-ARF6WT were treated with 5 mM CaCl2, for 10 minutes, fixed and stained for F-actin (red). The micrographs are representative of three independent experiments. Arrowheads indicate cells exhibiting ruffles and the arrow indicates the cell that did not ruffle. Bars, 10 µm. Right panels are a magnified view of inserts. (B) Quantification of CaSR-induced PM ruffling in cells presented in (A). *P<=0.001. NS, no significant difference (P>0.05, n=3). (C) Endogenous ARF6 is targeted to PM ruffles. CaSR-HEK cells were challenged with 5 mM CaCl2, for 10 minutes, fixed and stained for endogenous ARF6 (using a rabbit anti-ARF6 Ab followed by a Cy2-coupled anti-rabbit Ab, green) and for F-actin (red). Ruffles are indicated by arrowheads. Colocalisation areas appear in yellow. Bars, 5 µm.





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