Fig. 5

CGN c.3560C > T is associated with Rac1 activation. A Western blot of Rac1 expression level in HT-29 cells with CGN c.3560 C > C or c.3560C > T. B Densitometric quantification of GTP-Rac1 expression levels. Value, mean ± SEM from analysis of three different clones. C Representative confocal images of GTP-Rac1 (green) and DAPI nuclei (N) (blue) staining from 38 human tumor samples, including colorectal, ovarian, and endometrial cancers. Ten tumor samples are from patients with germline CGN c.3560C > T heterozygous mutation and 28 tumor samples are from patients with CGN c.3560C > C. Scale bar: 50 μm. D Quantitative analysis of Rac1 intensity from confocal images of 38 tumor samples using the TissueFAXS System. E Western blot showing IQGAP1, GTP-Rac1, and Rac1 protein levels. Two different IQGAP1-delivered siRNA duplexes or scramble siRNAs were treated for 24 hours followed 48 hours with or without 100 ng/ml EGF stimulus. Cell lysates were then harvested for blotting. Data are presented as the mean ± SEM. t-test for statistical significance. *p < 0.05, **p < 0.01