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. 2018 Feb 28;9:360. doi: 10.3389/fmicb.2018.00360

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Copyright © 2018 Bonfim-Melo, Ferreira and Mortara.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Rac1, Cdc42 and RhoA are recruited to actin-rich cup-like structures induced by EAs. HeLa cells were transfected with Cdc42, Rac1 (GFP tagged) or RhoA (c-myc tagged) and interactions with EAs were evaluated by live confocal microscopy. (A) Frames from time-lapse movies showing the moments of initial contact, beginning of actin recruitment and EA (arrowheads) internalization with recruitment and colocalization of Rac1-GFP (green) and LifeAct-RFP (red). (B) Frames from time-lapse movies showing Cdc42-GFP (green) recruited along with LifeAct-RFP (red) to EA (arrowheads) invasion sites during internalization. (C) Recruitment and colocalization of RhoA-c-myc (green) with actin (red) at EA invasion sites (arrowheads) was observed in fixed cells stained with phalloidin-TRITC. Nuclei and kinetoplasts were stained with Hoechst (A,B) or DAPI (C). Bar = 20 μm.