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Finally, immunostaining of whole kidney or western analysis of renal cortex Adenylyl cyclase or medulla did not reveal clear staining or bands for AC3 despite trying several different commercially available AC3 antibodies with varied antigen retrieval protocols (the latter for staining); this is a relatively common problem with detecting AC isoforms which have a very high degree of amino acid sequence homology between one another. Hence, to the best of our ability, and largely based on the DNA recombination data, it appears that AC3 was effectively targeted in the nephron. Figure 3 Relative mRNA expression of AC3 in the inner medulla from nephron AC3 KO mice and the controls. *P?http://www.selleckchem.com/products/tenofovir-alafenamide-gs-7340.html night time values between genotypes. Figure 4 Effect of nephron AC3 KO on systolic BP (A), diastolic BP (B), and heart rate (C) during normal (0.3%), high (4%), or low (0.01%) Na+ diets (n?=?5 mice/group). Nephron AC3 KO and control littermates were placed on a normal Na+ diet for 6?days, switched to a high salt diet for 7?days, followed BIBF-1120 by a low salt diet for 7?days. Urine volume and urinary Na+ excretion were similar between nephron AC3 KO and the controls on any of the days during different levels of Na+ intake (Fig.?(Fig.5).5). Food intake and water intake were similar between the two groups during normal, high, and low Na+ intake. Figure 5 Effect of nephron AC3 KO on urine volume (A) and urinary Na+ excretion (B) during normal (0.3%), high (4%), or low (0.01%) Na+ diets (n?=?9�C10 mice/group). Effects of nephron AC3 KO on plasma renin concentration As compared to a normal diet, PRC was increased in both nephron AC3 KO mice and the controls fed a low Na+ diet. There was no difference in PRC between the two genotypes during normal, high, or low Na+ feeding (Fig.?(Fig.66). Figure 6 Effect of nephron AC3 KO on plasma renin concentration during normal, high, and low Na+ intake (n?=?9�C10 mice/group). Effects of nephron AC3 KO on water handling Nephron AC3 KO and control mice were placed on a gel diet with free water access for 1?day followed by water restriction for 2?days. Water intake, urine volume, and urine osmolality were similar between nephron AC3 KO mice and the controls during normal water intake. There were no differences in urine volume and urine osmolality between the two genotypes during 2-day water restriction (Fig.?(Fig.77).