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Fig. 7. Effect of N17 Ras expression on LTD4- and EGF-induced Erk-1/2 activation. The cells were cotransfected with a vector expressing dominant-negative N17 Ras along with an empty EGFP vector and then stimulated with LTD4 (80 nM for 1 minute), EGF (100 ng/ml for 5 minutes) or not at all. The cells were then fixed, permeabilised and stained for phospho-Erk-1/2 as described in the Materials and Methods. The top three panels show from the left to the right: EGFP expression, phospho-Erk-1/2 staining and an overlay image of non-stimulated control cells. The middle three panels and the bottom three panels show the same analysis from cells stimulated with LTD4 and EGF, respectively. The results illustrated are representative of three separate experiments.





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