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. 2012 Jun 18;7(6):e37317. doi: 10.1371/journal.pone.0037317

Figure 5. Cdc42 loss does not impair transformation by the oncoprotein, c-Myc.

Figure 5

(a) p53-immortalized cells were infected with a retrovirus expressing the oncoprotein, c-Myc. c-Myc overexpression was verified by western blot analysis. (b) Pull-down assays were performed with GST-PAK1 to determine the relative abundance of activated Cdc42 in HRasV12 vs. c-Myc transformed cells. (c) Lysates from c-Myc transformed cells infected with Ad-GFP or Ad-GFP-Cre were immunoblotted for Cdc42. (d) Cultured c-Myc transformed cells were visualized 4 days following cdc42 deletion by bright field microscopy (upper 2 panels). Cells were fixed and stained with rhodamine-phalloidin and DAPI to visualize actin structures (lower 2 panels). (e) An equal number of c-Myc transformed Cdc42 proficient and deficient cells were seeded. Cells were counted every three days by trypan blue exclusion. Curves represent at least two independent experiments performed in triplicate. (f) Cdc42-proficient and –deficient c-Myc cells were grown in a 0.3% agarose solution. Experiments were performed in triplicate and 9 independent fields were counted for colonies. ns = not significant.