spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 1


Fig. 1. Membrane cholesterol and Cav-3DGV inhibit PDGF-induced DNA synthesis in fibroblasts. (A) CD and CO inhibit PDGF-induced DNA synthesis. Top panel: discontinuous stimulation for PDGF-induced DNA synthesis. Bottom panel: an example (left) and the statistical analysis (right) of the inhibitory effects of cholesterol depleting agents on PDGF-induced BrdU incorporation. Quiescent NIH 3T3 cells grown on coverslips were treated or not (NS) with CD (10 mM) or CO (0.5 U/ml) as indicated, rinsed and stimulated with PDGF (25 ng/ml) in the presence of BrdU. In some cases, cells have been incubated with CD in the presence of 25 µM soluble cholesterol before stimulation with PDGF (CD+Cholesterol). (B) Caveolin-1-GFP (Cav-1-GFP) and caveolin-3DGV (Cav-3DGV) inhibit mitogenesis induced by continuous PDGF stimulation. Top panel: continuous stimulation for PDGF-induced DNA synthesis. Bottom panel: an example (left) and the statistical analysis (right) of the inhibitory effect of indicated caveolin constructs on PDGF-induced BrdU incorporation. Cells transfected with the indicated constructs were made quiescent and stimulated for 18 hours with PDGF with or without soluble cholesterol (20 µM) as indicated and in the presence of BrdU. GFP was co-transfected with Cav-3DGV to visualise transfected cells. An example of BrdU incorporation obtained from Cav-3DGV transfected cells is indicated by arrows. (C) PDGF-induced DNA synthesis is reduced in cells grown in LPDS. NIH 3T3 cells were grown in 5% fetal calf serum (standard) or 5% LPDS in the presence or absence of soluble cholesterol (25 µM) as indicated for 30 hours and serum-starved for 24 hours before PDGF stimulation. An example (left) and the statistical analysis (right) of the inhibitory effects of chronically depleting cholesterol on PDGF-induced BrdU incorporation. Cells were fixed and BrdU incorporation was analysed by immunofluorescence as described in the Materials and Methods. The means and the s.d. from three to five independent experiments are shown.





Right arrow Return to article