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Exposure to the hormones triggers a rapid remodeling of the actin cytoskeleton and the formation of membrane ruffles required for cell movement, which are dependent on the rapid phosphorylation of the actin-regulatory protein moesin.
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Lamellipodia and membrane ruffles require Rac1; stress fibres and focal adhesions require RhoA; and filopodia require Cdc42.
Since membrane-ruffling requires the WRC (Hänisch et al., 2010) and our data show that ARNO specifically promotes this pathogen-induced ruffling and invasion, we used fluorescence microscopy to examine ARNO localization in HeLa cells expressing CFPARNO ARNOO) infected with WT or ΔE/E2 Salmonella.
P4.1 remained at membrane ruffles irrespective of the presence or absence of SipA (Figures S20).
In parallel, cell membrane ruffles and pseudopodia were formed at sites enriched in actin (Fig. 2A).
Subsequently, circular cup-shaped membrane ruffles closed into intracellular macropinosomes (Movie S1), as previously reported [29] [31].
Active membrane ruffles similar to those found in GFP-sec13 cells were not obvious.
Similarly Arg kinase showed co-localization with Unc119 at the membrane ruffles (Figure S3).
IQGAP1 colocalises with actin in lamellipodia and membrane ruffles [ 14].
Movie S4. 3D Visualization of MGC Membrane Ruffles, Related to Figure 6 3D visualization of MGC membrane ruffles as shown in Figure 6G.
Grp78 overexpressing cells formed obvious transparent membrane ruffles at leading edge, while the membrane ruffles was not obvious in mock-transfected cells (Fig. 3D).
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