1. Reduced expression and phosphorylation of NCC in SPAK243A/243A mice.
Reduced expression and phosphorylation of NCC in SPAK243A/243A mice. A. Expression and phosphorylation of NCC in wild type and knock‐in mouse kidney. Kidney extract derived from the indicated mice were subjected to immunoblot analysis with the specified NCC antibodies. Each sample is derived from a separate littermate animal. Band intensities were quantified using Li‐Cor Odyssey and the results are presented relative to the wild type (100%). This study has been repeated over six times each with kidney extracts derived from different animals on each occasion with consistent results. B. Expression and phosphorylation of NCC in wild type and knockin microdissected kidney tubules. Kidney tissue was microdissected from wild type (WT) or SPAK243A243A (KI) littermate mice. Tubular segments isolated are proximal tubule (PCT), cortical portions of the thick ascending limb (CTAL), distal convoluted tubule (DCT), connecting tubule (CNT) and cortical and outer medullary portions of the collecting duct (CCD). These were subjected to immunoblotting analysis with the indicated antibodies. C. Immunohistochemical analysis of the expression and phosphorylation of NCC in wild type and knock‐in mouse kidney. Immunohistochemical staining of whole kidney sections derived from the indicated mice probed with NCC total and phospho (Thr58) antibodies and visualized at a magnification of ×40.
Fatema H. Rafiqi et al. EMBO Mol Med. 2010;2:63-75
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