Figure 5.

Competitive binding confocal microscopy experiments with renal proximal tubules (RPT) isolated from normal rats (N=4 for each experiment). The experiment was repeated 4 separate times with new fresh kidneys. Within each experiment, each condition is represented by an average of 3 separate wells in a 96-well plate. Panel A. Effects of rostafuroxin (RF) binding on bodipy-oubain (B-OUA) binding. () Control: Opti-MEM. () B-OUA: 2 μM. () B-OUA + RF (0.5 μM). () B-OUA + RF (1.0 μM). () B-OUA + RF (10 μM). Control fluorescence values represent background. All other results are reported as B-OUA fluorescence intensity above background. Panel B. Effects of RF binding on biotinylated cGMP (BIO-cGMP). () Control: Opti-MEM. () BIO-cGMP: 2 μM. () BIO-cGMP + RF (10 μM). Control fluorescence values represent background. All other results are reported as BIO-cGMP fluorescence intensity above background. Data represent mean ± 1 SE. Scale bars represent 10 μm. Statistical significance was determined by using the repeated measures analysis with an unstructured covariance matrix in SAS PROC MIXED program. The ANOVA with permutation P value was based on 2,000 permutations of group assignment to individual N values and a repeated measures analysis with an unstructured covariance matrix.