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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: Cell Signal. 2011 Mar 16;23(8):1291–1298. doi: 10.1016/j.cellsig.2011.03.010

Figure 2. Quantification of cell ruffling and PLD2/Grb2 mutants.

Figure 2

F-actin-rich membrane protrusions in RAW/LR5 (A) or COS-7 cells (B) overexpressing YFP-PLD2, CFP-Grb2 or both proteins. PLD2 and Grb2 were visualized in the YFP and CFP channels, respectively. (C) Schemes representing the domains of both PLD2 and Rac2 and the location of the point mutants used in this study. (D) Effect of several mutants of PLD2 and Grb2 on cell ruffling quantitation in COS-7 cells. In A, B and D, visualization of cell ruffling was aided by Alexa-568-phalloidin staining. The number of protrusions was quantified using a scoring scale from 0 to 3 with 0 indicating no protrusion, 1 indicating protrusions in one area of the cell, 2 indicating protrusions in two distinct areas of the cell and 3 indicating protrusions in more than two distinct areas of the cell. The ruffling index was calculated by determining the average of the ruffling scores of at least 50 cells. (*) denotes statistical significance (p<0.05 by ANOVA) between Mock controls and means of indicated data points. (#) in panel (D) indicates an inhibition of the mutants over the PLD2-wild type transfections.