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. 2006 Nov 6;175(3):453–463. doi: 10.1083/jcb.200605144

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Copyright © 2006, The Rockefeller University Press

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Figure 7. — SopB-dependent Salmonella-induced actin remodeling requires RhoG. (A) Levels of Rac1 or RhoG in Henle-407 cells transfected with RNAi constructs that were directed to these GTPases measured by quantitative real-time RT-PCR. Values were standardized relative to the levels of Cdc42 mRNA, which are not affected by these constructs. (B and C) Intestinal Henle-407 cells were cotransfected with a plasmid expressing GFP (to detect transfected cells) along with RNAi constructs directed to Cdc42, Rac1, or RhoG or plasmids expressing Cdc42^N17, Rac1^N17, and vector control as indicated. Transfected cells were infected with wild-type S. typhimurium for 20 min (B) or its isogenic ΔsopE/ΔsopE2 (which induces actin remodeling solely through SopB) mutant for 40 min (C). After infection, cells were stained with rhodamine-phalloidin, and the percentage of transfected cells undergoing membrane ruffling as a consequence of bacterial infection was enumerated by fluorescence microscopy. Numbers indicate the percentage of transfected cells undergoing Salmonella-mediated actin remodeling (ruffling) and have been standardized considering the number of transfected cells undergoing membrane ruffling in control transfections to be 100%. The results represent the mean ± SD (error bars) of three independent experiments. At least 200 cells were counted for each independent transfection.