DOI: 10.5061/dryad.q1j50
title: Data from: A RhoG-mediated signaling pathway that modulates invadopodia dynamics in breast cancer cells

description: One of the hallmarks of cancer is the ability of tumor cells to invade
 surrounding tissues and metastasize. During metastasis, cancer cells
 degrade the extracellular matrix, which acts as a physical barrier, by
 developing a specialized actin-rich membrane protrusion structure called
 invadopodia. The formation of invadopodia is regulated by Rho GTPases, a
 family of proteins that regulates the actin cytoskeleton. Here, we
 describe a novel role for RhoG in the regulation of invadopodia
 disassembly in human breast cancer cells. Our results show that RhoG and
 Rac1 have independent and opposite roles in the regulation of invadopodia
 dynamics. We also show that SGEF is the exchange factor responsible for
 the activation of RhoG during invadopodia disassembly. When the expression
 of either RhoG or SGEF is silenced, invadopodia are more stable and have a
 longer lifetime than in control cells. Our findings also demonstrate that
 RhoG and SGEF modulate the phosphorylation of paxillin, which plays a key
 role during invadopodia disassembly. In summary, we have identified a
 novel signaling pathway involving SGEF, RhoG, and paxillin
 phosphorylation, which functions in the regulation of invadopodia
 disassembly in breast cancer cells.
description: Supplementary Movie 1.
 Invadopodia formation in CTRL SUM159 cells in response to
 PDBu.SUM159 cells expressing
 a CTRL shRNA were infected with mCherry-cortactin adenovirus and plated on
 glass-bottom coverslips 24 h after infection. Cells were imaged at 37°C in
 a CO2 and humidity controlled environment. At indicated times cells were
 stimulated to form invadopodia with PDBu. Images were captured at 15 sec
 intervals for the duration of the movie.Supp Movie 1-non targeting
 360x480_15fps.movSupplementary Movie 2.
 Invadopodia formation in Rac1 KO cells.Rac1 KO SUM159 cells
 were infected with adenovirus encoding mCherry-cortactin as in Supp Movie
 1. At indicated times cells were stimulated to form invadopodia with PDBu.
 Images were captured at 15 sec intervals for the duration of the
 movie.Supp
 Movie 2-Rac KO-360x480.movSupplementary Movie 3.
 Invadopodia formation in Rac1 KO/rescue cells.Rac1 KO SUM159 cells
 were co-infected with adenovirus encoding mCherry-cortactin and mycRac1.
 At indicated times cells were stimulated to form invadopodia with PDBu.
 Images were captured at 15 sec intervals for the duration of the
 movie.Supp
 Movie 3-Rac KO-
 Rescue_360x480.movSupplementary Movie 4.
 Invadopodia in RhoG KD SUM159 cells.SUM159 cells stably
 expressing a RhoG targeting shRNA were infected with mCherry-cortactin
 adenovirus as in Supp Movie 1. At indicated times cells were stimulated to
 form invadopodia with PDBu. Images were captured at 15 sec intervals for
 the duration of the movie.Supp Movie 4_RhoG KD_480x360
 37min.movSupplementary Movie 5.
 Invadopodia formation in SGEF KD cells.SUM159 cells stably
 expressing SGEF targeting shRNA were infected with mCherry-cortactin
 adenovirus as in Supp Movie 1. At indicated times cells were stimulated to
 form invadopodia with PDBu. Images were captured at 15 sec intervals for
 the duration of the movie.Supp Movie 5
 -SGEF-KD-360x480.movSupplementary Movie 6.
 Invadopodia formation in RhoG KD/rescue cells.RhoG KD SUM159 cells
 were co-infected with adenoviruses encoding mCherry-cortactin and shRNA
 resistant mycRhoG. At indicated times cells were stimulated to form
 invadopodia with PDBu. Images were captured at 15 sec intervals for the
 duration of the movie.Supp Movie
 6-RhoG-rescue_480x360.movSupplementary Movie 7.
 Invadopodia formation in SGEF KD/rescue cells.SGEF KD SUM159 cells
 were co-infected with adenovirus encoding mCherry-cortactin and shRNA
 resistant mycSGEF. At indicated times cells were stimulated to form
 invadopodia with PDBu. Images were captured at 15 sec intervals for the
 duration of the movie.Supp Movie 7_SGEF-rescue
 480x360.mov