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Figure 4


Fig. 4. Laminin-1 induces clustering of β1 integrin in lipid rafts. (A) PC12 cells were treated with 100 ng/ml of NGF and/or 5 µg/ml of laminin-1. Cells were lysed in 1% Triton X-100 and fractionated on discontinuous sucrose gradients. Ten fractions were collected from top to bottom. Each fraction was subjected to immunoblotting using the indicated antibodies (lanes 1-10). Flotillin-1, a raft marker, was detected in fractions 2 and 3. (B) DRG cells, labeled with BODIPY-GM1 (green) and Alexa-Fluor-546-conjugated anti-β1 integrin antibody (red), were treated with 5 µg/ml of laminin-1. DRG cells were analyzed using confocal laser microscopy. Arrowheads indicate clustering and colocalization of BODIPY-GM1 and β1 integrin. Inset shows an enlarged image of the boxed area. Bar, 5 µm.





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