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. Author manuscript; available in PMC: 2014 Mar 1.

Published in final edited form as: Kidney Int. 2013 May 1;84(3):468–481. doi: 10.1038/ki.2013.149

(A) EpoR mRNA expression in the rat kidney or microdissected glomeruli and renal tubules and from normal adult rats at age of 3 months old by RT-PCT. Total RNA was extracted, and complimentary DNA (cDNA) generated with Oligo dT. Specific target genes were examined by PCR with rat specific primers (shown in method section). AQP2: aquaporin-2; CCD: cortical collecting duct; DCT: distal convaluted tubules; Glo: glomeruli; IMCD: inner medullary collecting duct; K: Rat kidney tissue containing cortex and medullar; NaPi-2a: Na-Pi dependent cotransporter-2a; NKCC2: Na-K-2Cl cotransporter; PT: proximal tubules; TAL: thick ascending limb; K-RT: kidney sample with reverse transcriptase omitted; (B) EpoR protein expression in BaF3 cell transfected with HA-tagged mouse EpoR or empty vector. Total lysates from native BaF3 and BaF3-HA-EpoR cells were subjected to immunoprecipitation by HA-Resin followed by immunobloting for EpoR with several anti-EpoR antibodies: A82, HA, M-20, Fab 6. Asterisks depict the specific HA-EpoR band. (C) Total proteins were extracted from human and rodent kidneys and murine FLC and Ter119-cells, and subjected to immunobloting EpoR with several antibodies against EpoR. A82 is monoclonal rabbit antibody kindly provided by Dr. Steve Elliot; M-20 is polyclonal antibody purchased from Santa Cruz Biotech. Fab 6 is a synthetic human Fab against human EpoR isolated from a phage-displayed library. Asterisks indicate multiple forms of glycosylated EpoR or EpoR fragments. Hu: human; Mu: mouse; FLC: fetal liver cells at E13.5

(A) EpoR mRNA expression in the rat kidney or microdissected glomeruli and renal tubules and from normal adult rats at age of 3 months old by RT-PCT. Total RNA was extracted, and complimentary DNA (cDNA) generated with Oligo dT. Specific target genes were examined by PCR with rat specific primers (shown in method section). AQP2: aquaporin-2; CCD: cortical collecting duct; DCT: distal convaluted tubules; Glo: glomeruli; IMCD: inner medullary collecting duct; K: Rat kidney tissue containing cortex and medullar; NaPi-2a: Na-Pi dependent cotransporter-2a; NKCC2: Na-K-2Cl cotransporter; PT: proximal tubules; TAL: thick ascending limb; K-RT: kidney sample with reverse transcriptase omitted; (B) EpoR protein expression in BaF3 cell transfected with HA-tagged mouse EpoR or empty vector. Total lysates from native BaF3 and BaF3-HA-EpoR cells were subjected to immunoprecipitation by HA-Resin followed by immunobloting for EpoR with several anti-EpoR antibodies: A82, HA, M-20, Fab 6. Asterisks depict the specific HA-EpoR band. (C) Total proteins were extracted from human and rodent kidneys and murine FLC and Ter119-cells, and subjected to immunobloting EpoR with several antibodies against EpoR. A82 is monoclonal rabbit antibody kindly provided by Dr. Steve Elliot; M-20 is polyclonal antibody purchased from Santa Cruz Biotech. Fab 6 is a synthetic human Fab against human EpoR isolated from a phage-displayed library. Asterisks indicate multiple forms of glycosylated EpoR or EpoR fragments. Hu: human; Mu: mouse; FLC: fetal liver cells at E13.5