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1.
Figure 4

Figure 4. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

Association of CAV1 with GRK4 in nRPTCs and uRPTCs. CAV1 and GRK4 association was measured as the amount of immunoreactive GRK4 by Western blot of immuno-precipitated CAV1 (shown in top insert) using a monoclonal antibody. Immunoreactive GRK4 associated with CAV1 was decreased in uRPTCs vs nRPTCs (*P<0.05 vs nRPTC, t test; n=4 per group).

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.
2.
Figure 6

Figure 6. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

AP2-mediated internalization of NaKATPase. Immuno-precipitated AP2 followed by NaKATPase dot blot is expressed as percentage of VEH in nRPTCs (□) and uRPTCs (). FEN (1 μmol/L for 30 minutes) stimulated the AP2-mediated NaKATPase internalization in nRPTCs but not in uRPTCs. The FEN effect in nRPTCs was blocked by βMCD or CAV1 siRNA (*P<0.05 βMCD FEN vs VEH FEN in nRPTC, n=6 per group; #P<0.05 CAV1 siRNA FEN vs SCR FEN in nRPTCs, n=6 per group).

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.
3.
Figure 5

Figure 5. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

Effect of βMCD on the plasma membrane CAV1 expression in nRPTCs and uRPTCs. βMCD (2 mmol/L; 1 hour), a cholesterol-reducing agent, decreased CAV1 expression in nRPTCs but not in uRPTCs. Membrane CAV1 was reduced in nRPTCs by βMCD to the same level of CAV1 expression in uRPTCs (*P<0.05 vs nRPTC PBS VEH, ANOVA, Holm-Sidak test; n=6 per group).

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.
4.
Figure 3

Figure 3. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

Western blot analysis of total cellular expression of CAV1 in nRPTCs treated with VEH, FEN, or ANG II. FEN (1 μmol/L; 4 hours) increased whereas ANG II (10 nM; 4 hours) decreased the total cell expression of CAV1 (*P<0.05 vs others, ANOVA, Holm-Sidak test; n=8 per group). β-Tubulin (β-Tub) shows even loading of proteins.

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.
5.
Figure 2

Figure 2. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

Western blot analysis showing the effect of DMSO VEH or FEN (1 μmol/L for 30 minutes) on plasma membrane expression of D1R, GRK4, and CAV1 in nRPTCs and uRPTCs. The results from representative Western blots are shown above each bar graph (*P<0.05 vs nRPTC VEH; n=4 for each graph). A, Increase in D1R plasma membrane expression after FEN (1 μmol/L for 30 minutes) in nRPTCs but not in uRPTCs. B, Decrease in GRK4 plasma membrane expression after FEN (1 μmol/L for 30 minutes) in nRPTCs but not in uRPTCs. The basal expression of plasma membrane GRK4 was lower in uRPTCs than in nRPTCs. C, Increase in CAV1 plasma membrane expression after FEN stimulation seen in nRPTCs but not in uRPTCs. The basal expression of CAV1 was lower in uRPTCs vs nRPTCs (*P<0.05 vs VEH, nRPTCs). D, Loading controls of β-tubulin (β-Tub) and Ponceau S staining.

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.
6.
Figure 1

Figure 1. From: Caveolin-1 and Dopamine-Mediated Internalization of NaKATPase in Human Renal Proximal Tubule Cells.

Comparison of FEN-stimulated coupling efficiency with adenylyl cyclase in nRPTCs and uRPTCs. A, cAMP accumulation using ELISA, in nRPTCs or uRPTCs after stimulation for 30 minutes with either DMSO vehicle control (CON), 1 μmol/L of FEN, or FEN with 10 μmol/L of LE300, a D1-like receptor antagonist (FEN LE300; *P<0.05 vs others; n=6 per group). B, Intracellular cAMP accumulation in real time using a cAMP fluorescence resonance energy transfer biosensor, ICUE3, measured in nRPTCs and uRPTCs. FEN (1 μmol/L; marked by an arrow at 1 minute) increased intracellular cAMP in nRPTCs vs uRPTCs. Data are from 3 separate wells of 10 cells per well, imaged simultaneously. Graph is representative of 3 replicate experiments.

John J. Gildea, et al. Hypertension. ;54(5):1070-1076.

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