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Fig. 6. Invasion and signal transduction. (A,B) Co-transfection of CapG-EGFP with RhoAN19 or Cdc42N17 (lanes 4-5), but not with Rac1N17 (lane 3), inhibits CapG-induced collagen type I invasion of HEK293T cells (A) (mean ± s.e.m., n=3, EV = empty vector) or HCT8/E-11 colon adenocarcinoma cells (B). (C) Co-expression of RasGAP (lane 5), p85{Delta}iSH2N (`PI3KDN', lane 6) or dominant-negative MEK (lane 7) with CapG-EGFP blocks invasion. RasV12C40 shows no inhibition of invasion (lane 10) in contrast to RasV12S35 (lane 8) and RasV12E38 (lane 9). Values are representative of three independent experiments (mean ± s.e.m.). (D) Upper: western blot on transfected HEK293T c ells. Numbers correspond to the numbers in the histogram in (C). Lower: tubulin western blot = control. (E) Western blots showing expression of RasV12, RasGAP, p85{Delta}iSH2N, RasV12E38, RasV12C40, RasV12S35 and MEKDN in cells co-transfected with CapG-EGFP. (F) Collagen invasion triggered by CapG-EGFP is counteracted by MMP inhibitors (mean ± s.e.m., n=3): aprotinin (10 µg/ml), galardin (10 µM) and the corresponding negative control (10 µM), TIMP-1 (500 ng/ml). CTTHWGFTLC = decapeptide (50 µM).





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