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Figure 6


Fig. 6. PDGFRβ remains stable in PDGF-BB-induced and -internalized LRP1 complexes in MT1-MMP-expressing cells. (A) Control (+/+) and MT1-MMP-null (–/–) VSMCs on collagen were treated with PDGF-BB for 30 minutes and the subcellular localization of LRP1 and PDGFRβ analyzed by immunofluorescence. Note the intracellular colocalization of these receptors in PDGF-BB-treated wild-type VSMCs. (B) VSMCs were surface biotinylated and shifted to 37°C for 30 minutes in the presence of PDGF-BB to allow PDGFRβ internalization. The biotinylated PDGFRβ remaining on the cell surface was left intact (biotinylated) or removed by reduction (internalized). Total biotinylated and internalized receptors were then detected in PDGFRβ immunoprecipitates by streptavidin conjugate. Total and ubiquitylated PDGFRβ in precipitates was detected by immunoblotting. (C) VSMCs were stimulated with PDGF-BB for the indicated periods of time (minutes) and lysed for analysis. Total PDGFRβ protein levels were assessed by immunoblotting. Relative mobilities of the molecular-mass markers are indicated in kDa.





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