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. 2011 Feb;13(2):98–107. doi: 10.1593/neo.101088

Figure 2.

Figure 2

Figure 2

H-Ras HVRs are critical for the activation of the Rac-MKK3/6-p38 pathway. (A) For Rac1 activity, cell lysates were incubated with GST-PBD fusion protein, and the bound active Rac1-GTPmolecules were analyzed by immunoblot analysis using an anti-Rac1 antibody. The levels of activated MKK3/6, p38 in Ras chimera, and mutant cells were determined by immunoblot analysis of whole cell lysates. Band intensities were quantitated and plotted. *,**Statistically different at P < .05 and P < .01, respectively. (B) Cells were double labeled for H-Ras or N-Ras (green) and pRac1 (red). Immunocytochemical colocalization of H-Ras or N-Ras and flotillin-1 was observed by confocal microscopy. (C) H-Ras MCF10A cells were transfected with control siRNA or siRNA H-Ras. Protein level of H-Ras was detected by immunoblot analysis. β-Actin was used as a loading control. Rac1 activity (Rac1-GTP) was measured as described in Materials and Methods.