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Figure 2


Fig. 2. Membrane-cholesterol depletion does not affect PDGF-induced receptor, Ras and ERK activation. (A) CD does not affect PDGF receptor activation. NIH 3T3 cells were serum starved, treated with CD, rinsed and stimulated with PDGF for 10 minutes as indicated. PDGFRß was immunoprecipitated from indicated lysate with {alpha}PR4 followed by in vitro kinase assay (left panel). The position of [32P]PDGFRß is shown. Total cell lysates treated as indicated were directly subjected to western blotting with anti-phosphotyrosine antibody (4G10) (right panel). The location of molecular markers (Mr) is also shown. (B) CD does not affect PDGF-induced Ras (left panel) and ERK (right panel) activation. Lysates were either directly subjected to western blotting with anti-Ras (total Ras), anti-ERK (ERK) and anti-phospho ERK (pERKs) antibodies or incubated with Sepharose-bound GST-RalGDS binding domain and western blotting with anti-Ras antibodies (Ras-GTP) as indicated.





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