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Fig. 10. MDCK P21R (MDCK/P21R) cells display increased expression levels of the endogenous EGFR. (A) Ectopic expression of P2Y1R increases EGFR levels. Immunoblots of total cell extracts, densitometrically quantified in four independent experiments and normalized with respect to actin show a threefold increased level of EGFR in MDCK P2Y1R cells. (B) Ectopic expression of P2Y1R increases EGFR biosynthesis. Newly synthesized EGFR was immunoprecipitated from cells metabolically labeled for 2 hours with [35S]Met/Cys (200 µCi/ml). Graph shows twofold increase in synthesis in MDCK P2Y1R cells (n=3). For A and B, values are the means ± s.e.m. ***P<0.001; paired t-test compared with wild-type cells. (C) Increased EGFR expression in MDCK P2Y1R cells depends on EGFR and PKC activity. Cells were treated with either 100 nM AG1478 or 10 nM Ro318220 for 6-18 hours before and during the metabolic labeling. Graph show the densitometric quantification of three independent experiments. Values are the means ± s.e.m. *P<0.05, **P<0.01; Dunnett's test compared with untreated cells.
