Dopamine 5 receptor mediates Ang II type 1 receptor degradation via a ubiquitin-proteasome pathway in mice and human cells
J. Clin. Invest. Hewang Li, et al. 118:2180 doi:10.1172/JCI33637 [Go to this article.]

Figure 3
Effect of D₅R stimulation on the expression of the glycosylated form of the AT₁R protein. (A) AT₁R/D₅R HEK293 cells were incubated with Fen (1 μM) for the indicated times; total cell lysates were electrophoretically separated, transferred onto nitrocellulose membranes, and immunoblotted with anti-GFP (band a, ~100–120 kDa; band b, ~65–85 kDa) to detect AT₁R protein expression. Immunoblotting for actin was used to monitor the sample protein loading. n = 3. *P < 0.05 versus 0 min, ANOVA, Student-Newman-Keuls test. (B) Protein samples (50 μg) from AT₁R/D₅R HEK293 cells were incubated with 10 mU PNGase F at 37°C for the indicated times, electrophoretically separated, transferred onto nitrocellulose membranes, and immunoblotted with anti-GFP antibody. Band S is sensitive to PNGase F and corresponds to the glycosylated form of the AT₁R (band a). Band R is resistant to PNGase F and corresponds to the nonglycosylated form of the AT₁R (band b). The ratio of band S intensity to band R intensity was determined by densitometry in 3 separate experiments. n = 3. *P < 0.05 versus 0 h, ANOVA, Student-Newman-Keuls test. Data are mean ± SEM.