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. 2014 Dec 1;7:95. doi: 10.3389/fnmol.2014.00095

FIGURE 1.

FIGURE 1

Generation of transgenic rats. (A) Structure of NSE-NPR-BΔKC transgenic construct using the neuron-specific enolase promoter (NSE) for neuron-specific expression. The flag/tag was used for immunodetection of the expressed protein. (B) Detection of a founder animal (8809, boxed lane) by Southern blotting. Native NPR-B is present in wild type animals (WT), and native NPR-B and transgene are present in the positive control and founder animal 8809. The transgene construct has a size of 950 bp, and native NPR-B of approximately 10 kbp. (C) Specific expression of NPR-BΔKC-Flag was demonstrated by RT-PCR. Negative control (NTC) and positive control (control + transgene) are shown for the RT-PCR showing NPR-BΔKC-Flag expression. PCR for GAPDH expression was used as loading control. M indicates a 100 bp DNA fragment ladder. (D) Specific expression of NPR-BΔKC-Flag as shown by Western blotting using an anti-flag antibody using extracts from brain, heart, lung, aorta, and kidney. GAPDH expression is shown as a loading control. (E,F) NPR-BΔKC overexpression significantly reduced CNP-dependent, but not ANP-dependent cGMP production in brain membrane preparations in transgene animals (TG) compared to wild-type animals (WT). *p < 0.05, **p < 0.01 vs. wild-type (n = 6 per group).