Fig. 7. Analysis of cell-cell adhesion in invasive T98G cells. (A) Matrigel invasion assay using as chemoattractants either serum-free medium containing 0.1% BSA (basal) or conditioned media from the same cells cultured in Matrigel-coated Petri dish (stimulated). The data are the mean number of cells per field±s.d. of triplicate samples from a representative experiment. (B) Western blot analysis of cadherin, ß-catenin and LIN-7 total expression in T98G cells cultured on poly-L-lysine (lane 1) or Matrigel (lane 2). The same amounts of total cell homogenates were loaded onto 11% SDS-PAGE and immunoblotted with the indicated antibodies. Molecular weight standards expressed in kDa are indicated on the left. (C) Brightfield and (D) confocal laser microscopy of T98G cells cultured in Matrigel. The cells (2x10⁵ cells/35 mm diameter dish) were seeded onto Matrigel-coated glass coverslips and cultured for three days. After fixation in 4% paraformaldehyde, the cells were double-stained for ß-catenin and the indicated antibodies. The arrows indicate ß-catenin-enriched cell-cell contacts with disorganised filaments of F-actin, virtually no LIN-7 and the accumulation of phosphorylated proteins. Bar, 10 µm.

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