Functional differences in HAB and LAB NPSR1 variants. A, B, HEK-CRE-luc cells were transfected with rHAB or rLAB Npsr1 cDNA constructs carrying A(227,016)G (A) and mHAB or mLAB Npsr1 cDNA constructs carrying A(156,453)G (B); rs37572071 along with Gaussia vector and stimulated with 1 nmol/5 μl NPS at 40 h after transfection until assay at 48 h. C, NPSR1 genotyping using hypothalamic tissue from WT and NPSR1 KO mice. D, E, Semi-quantitative Western blots for total NPSR1 protein of WT and NPSR1 KO mice in hypothalamic paraventricular nucleus tissue with Abcam antibody ab92425 (43 kDa; D) and with Ab2 (72 kDa; Leonard and Ring, 2011; E). β-Tubulin served as a loading control for both Western blots. Data are shown as the mean ± SEM, and numbers in parentheses indicate the group size. **p < 0.01. The luciferase assay was performed independently, and each n was an average of three replicates.