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Figure 5


Fig. 5. Rac3 phenocopies RhoA in N1E-115 cells. (A) Cells were transfected with pcDNA3.1/Rac3, pcDNA3.1/RhoAG14V, pSuper/shRac3 or pSuper/shRhoA, together with 10:1 eGFP. Subsequently, the cells were fixed and stained with phalloidin to visualize F-actin. Bar, 25 µm. Note that both Rac3 and RhoA overexpression induce cell rounding, whereas the depletion of either protein leads to protrusion formation. (B) Morphologies were scored as flat, round or protrusion bearing and shown here in a bar graph. At least 200 cells from two independent experiments were counted. (C) Neurite-like protrusions induced by shRac3 are not responsive to LPA. N1E-115 cells were transfected with either shLuc or shRac3 together with 5:1 eGFP to visualize the transfected cells. Cells were either stimulated or not with 5 µM LPA for 15 minutes and subsequently photographed. (D) Quantification of the results shown in (C). Protrusions-bearing cells were counted in three independent experiments and represented in a bar graph.





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