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. 2005 Aug;25(16):6921–6936. doi: 10.1128/MCB.25.16.6921-6936.2005

FIG. 5.

FIG. 5.

The TGFβ1 effects depend on expression and activation of integrins α2 or α3. The cells were pretreated with 10 μg/ml anti-integrin α2 (P1E6), α3 (P1B5), or α5 (P1D6) antibodies, 30 min prior to the replating on Fn and a concomitant TGFβ1 treatment for 20 h. After the incubation, cell images were taken, and lysates were prepared and used in immunoblots using antibodies against the indicated molecules. Data shown are representative of two isolated experiments. (A) Functional blocking anti-integrin α2 or α3, but not α5, antibodies abolished the cell spreading. (B) Functional blocking of the integrins inhibited phosphorylation of the FA molecules. (C) Cells were transiently cotransfected with a control plasmid (C) or pSF2-integrin α2 or α3 with pCDNA3-GFP. One day after the transfection, cells were replated onto Fn (10 μg/ml)-precoated cover glasses in the absence of TGFβ1 treatment. After incubation for 20 h at 37°C and 5% CO2, cells were processed for actin staining with phalloidin-conjugated TRITC. Another set of cells in 60-mm culture dishes were harvested for lysates prior to performing immunoblotting using the indicated antibodies. BLK mAb, functional blocking monoclonal antibody.