JCI - Vasopressin-induced von Willebrand factor secretion from endothelial cells involves V2 receptors and cAMP

Vasopressin-induced von Willebrand factor secretion from endothelial cells involves V2 receptors and cAMP

Jocelyne E. Kaufmann, Alexander Oksche, Claes B. Wollheim, Gabriele Günther, Walter Rosenthal, Ulrich M. Vischer

J Clin Invest. 2000; 106(1):107–116 doi:10.1172/JCI9516

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Figure 6

Expression of V2R in HMVEC-L and human lung. (a) RT-PCR amplification from human kidney, HMVEC-L, HUVECs, and human lung polyA⁺. The expected 857-bp fragment (arrow) was amplified from V2R mRNA using primers V2Rc and V2Rd. The RT-PCR reaction was performed in absence or presence of RT. First lane: HindIII/EcoRI lambda marker. The gel was stained with ethidium bromide. (b) Southern blot analysis of the RT-PCR products amplified from kidney, lung, HUVECs, and HMVEC, using a probe derived from V2R cDNA. Negative control: RT-PCR reaction performed in absence of template. The arrow indicates V2R transcript (857 bp); the band visible just above represents genomic V2R DNA. Five times less of the RT-PCR reaction was loaded on the gel for both kidney and lung compared with HUVECs and HMVEC-L. (c) RT-PCR amplification of reversed transcribed polyA⁺ RNA from human lung from two different donors (lanes 3 and 4) using primers STV2 and V2RBamHI. The RT-PCR reaction was performed without (lanes 3 and 4) or with prior pretreatment of the samples with RNase-free DNaseI (lane 5). The size of the fragment expected to be amplified from V2R mRNA is 1,144 bp, as verified by amplification of plasmid DNA carrying the full-length coding region of the V2R (lane 2). Lane 1: RT-PCR amplification of plasmid DNA harboring the genomic DNA of the V2R (expected size: 1,611 bp). The gel was stained with ethidium bromide.