Journal: The FASEB Journal
Article Title: Urinary concentrating defect in mice lacking Epac1 or Epac2
Figure Lengend Snippet: AQP2 expression in Epac1 −/− and Epac2 −/− mice after water deprivation. A ) Representative Western blots from whole kidney lysates of Epac WT, Epac1 −/− , and Epac2 −/− mice subjected to 24 h of water deprivation probed with anti-AQP2 antibodies. AQP2 is present as upper duplet of glycosylated bands around 37 kDa and lower nonglycosylated band at 29 kDa. B ) Summary graph comparing total renal AQP2 expression (both glycosylated and nonglycosylated forms) in Epac WT, Epac1 −/− , and Epac2 −/− from Western blots similar to that shown in A . Intensity values were normalized to total signal of respective lines in Ponceau red staining. Number of mice in each group is indicated on each bar. C ) Representative confocal micrographs of cortical (top) and medullary (bottom) CDs in transverse cut kidney sections probed with anti-AQP2 (pseudocolor green) from Epac WT, Epac1 −/− , and Epac2 −/− mice subjected to 24 h of water deprivation. Nuclear DAPI staining is shown in pseudocolor blue. Number of mice in each group is indicated on each bar. * P
Article Snippet: We used the primary antibodies against anti-AQP2 (1:1000, AQP2-002; Alomone Labs, Jerusalem, Israel), anti–Na+ -K+ -Cl− cotransporter type 2 ( SLC12A1 ; NKCC2; rabbit polyclonal, 1:1000, ANT-072; Alomone Labs), anti–NHE-3, pS552 NHE-3 (mouse monoclonal, 1:100, sc-136368, sc-53962; Santa Cruz Biotechnology, Dallas, TX, USA), anti-Na+ /H+ exchanger regulatory factor isoform 1 (NHERF1; mouse monoclonal, 1:1000, sc-271552; Santa Cruz Biotechnology), anti-pT96 T101 NKCC2 (rabbit polyclonal, 1:2000, gift of B. Forbush, Yale University, New Haven, CT, USA), anti-pT53 thiazide-sensitive Na+ -Cl− cotransporter ( SLC12A3 ) (NCC; rabbit polyclonal, 1:1000; p1311-53; PhosphoSolutions, Aurora, CO, USA), anti–α subunit of epithelial Na+ ).
Techniques: Expressing, Mouse Assay, Western Blot, Staining