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Fig. 4. Effect of FRNK overexpression in GM7373 endothelial cells. (A) Parental GM7373 cells or the indicated transfectant clones were grown on tissue culture plates, lysed and immunoblotted with an antibody directed against the C terminus of FAK. (B) The different clones were allowed to adhere to plastic coated with 20 µg/ml of GST-Tat (black bars), FN (hatched bars) or BSA (white bars). At the end of incubation, cell adhesion was quantified. Each point is the mean ± s.e.m. of three determinations in duplicate. (C) Representative images showing parental and FRNK-transfected GM7373 cells adherent to GST-Tat or FN (magnification 100x). (D) The different clones were seeded on cell culture plates, incubated for 30 minutes with or without free GST-Tat (100 ng/ml), and immunoblotted with antibodies specific for FAK phospho-Tyr₃₉₇ residue. Alternatively, the different clones were allowed to adhere to plates coated with GST-Tat or PL and analysed for RhoA or pp60^src activation. No significant differences were found in the total/starting levels of FAK, pp60^src or RhoA (data not shown). The data are representative of two to three independent experiments that gave similar results.

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