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Fig. 4. CD9 ligation induces tyrosine phosphorylation of p46 Shc. (A) MKN-28 cells were serum starved for 24 hours and then stimulated with 50 µg ml^–1 ALB6 or isotype-matched mouse IgG1 for 10 minutes. The total cell lysates were immunoprecipitated with the indicated antibodies and the blots were analysed with PY20 (top). The same filters were then stripped and reprobed with the indicated antibodies to confirm equal loadings (bottom). Each figure shows one of four similar experiments. (B) MKN-28 cells were serum starved for 24 hours and subsequently cultured for 48 hours in the presence of 50 µg ml^–1 isotype-matched mouse IgG1, 50F11 or ALB6, and their proliferation was measured by a [³H]-thymidine incorporation assay. Data are represented as the means±SEM from six replications and are expressed as a percentage of the values found for isotype-matched mouse IgG1 (50 µg ml^–1) treatment cells (top). *P<0.01 vs control. MKN-28 cells were serum starved for 24 hours, and then stimulated with 50 µg ml^–1 isotype-matched mouse IgG1, 50F11 or ALB6 for 10 minutes. The total cell lysates were immunoprecipitated with anti-Shc antibody and the blots were analysed with PY20 (bottom). (C) MKN-28 cells were serum starved for 24 hours and were subsequently pretreated either 10 µM PP2 or 10 µM PP3 for 30 minutes, and then stimulated with or without 50 µg ml^–1 ALB6 for 10 minutes. The total cell lysates were immunoprecipitated with anti-Shc antibody and the blots were analysed with PY20.

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