Journal: American Journal of Physiology - Renal Physiology
Article Title: Differences in renal BMAL1 contribution to Na + homeostasis and blood pressure control in male and female mice
doi: 10.1152/ajprenal.00014.2020
Figure Lengend Snippet: Characterization of kidney-specific brain and muscle ARNT-like 1 (BMAL1) knockout (KS-BMAL1 KO) mice. A: model showing exon 8 of BMAL1 excision by Ksp-cadherin Cre. Solid gray arrows represent primers used for recombination PCR. *Band produced from the recombined floxed gene. B: recombination PCR assay showing the expected 570-bp product (*) in renal inner medullary tissue of KS-BMAL1 KO mice but not control (CNTL) mice. C: additional assays assessing nonrenal tissue revealing the lack of the KO band. D−I: representative images of BMAL1 protein expression in renal segments. D, F, and H: colabeling with BMAL1 (brown) in CNTL mice; E, G, and I: colabeling with BMAL1 (brown) in KS-BMAL1 KO mice. D and E: blue staining with Na+-K+-2Cl− cotransporter 2 (NKCC2) representing the cortical thick ascending limb (TAL). F and G: blue staining with phosphorylated Na+-Cl− cotransporter (p-NCC) to identify distal convoluted tubules (DCTs). H and I: blue staining with aquaporin 2 representing medullary collecting duct (CD) cells. Both PCR and immunohistochemistry images are from male tissue except for ovary tissue from female mice. Arrows indicate BMAL1-positive stained nuclei; arrowheads indicate nuclei with negative BMAL1 staining. J: immunoblots of renal inner medullary homogenates from male mice to demonstrate BMAL1 protein expression and total protein from Ponceau staining. The predicted size of BMAL1 band (78 kDa) is noted. K: quantification determined by normalizing BMAL1 values with total protein. PT, proximal tubule. Values are means ± SE; n = 4. Student’s t test was used to compare genotypes. Scale bar = 0.05 mm.
Article Snippet: Antibody information is shown in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antibody Target Manufacturer Catalog Number Reference Description Dilution Used RRID AQP2 Stressmarq SPC-503D 35 Rabbit, polyclonal IHC: 1:400 AB_10965071 BMAL1 Cell Signaling D2L7G 18 Rabbit, polyclonal IHC: 1:3,000; Western blot analysis: 1:1,000 AB_2728705 NKCC2 Stressmarq SPC-401D 19 Rabbit, polyclonal IHC: 1:10,000 AB_10640877 T53 p-NCC Previously characterized 30 Rabbit, polyclonal IHC: 1:1,000,000 Not applicable Open in a separate window Antibodies were used in this study as previously described ( 18 , 19 , 30 , 35 ).
Techniques: Knock-Out, Produced, Expressing, Staining, Immunohistochemistry, Western Blot
StressMarq is a verified supplier 
StressMarq manufactures this product 
