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Fig. 8. Activation and functions of the MAPK signaling pathway in crest cells cultured on LN-1 and E8 and E1' fragments. (A) Immunoblotting for phosphorylated Erk (upper panel) and for pan-ErK (lower panel) on lysates of crest cells cultured on LN-1, heat-denatured LN-1 (D-LN1), E8 and E1'. (B) Immunoblotting for phosphorylated Erk on lysates of cells cultured on LN-1 and on antibodies against 
1ß1. Note that, as described previously for chicken embryo fibroblasts (Fincham et al., 2000), only a single species of Erk immunoreactivity is detected in crest cells. (C-E) Immunofluorescence staining for phosphorylated Erk in crest cells cultured for 24 hours on LN-1, E8 and E1' at 10 µg/ml. All images were acquired with the same exposure duration. (F-H) Adhesion, spreading and migration on LN-1, E1' and E8 in the presence of the MEK inhibitor PD98059. In all assays, cells were incubated with 5 µg/ml PD98059 throughout the duration of the assay and, in the adhesion and spreading assays, cells were preincubated in suspension with the drug for 10 minutes prior to the assay. (I-K) Morphology and migration of cells over LN-1, E1' and E8 in the presence of 5 µg/ml PD98059. (L-N) Cell survival and morphology in the presence of PD98059. Cells were cultured for up to 24 hours on their substrate to allow complete spreading and the drug was added at 5 µg/ml for 4 hours.
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