Npr2-7A inactivates Npr2. A, Npr2 and Npr2-7A coimmunoprecipitate. The 293T cells were transfected with 1 μg GFP plasmid, 2 μg FLAG-mouse-Npr2-WT, 1 μg FLAG-mouse-Npr2-WT, and 1 μg HA-human-Npr2-7A or 2 μg HA-human-Npr2-7A. After 48 h, crude membranes were prepared and assayed for guanylyl cyclase activity. The remaining membranes were lysed, and then the Npr2 proteins were immunoprecipitated with the indicated antibodies. The proteins were separated by SDS-PAGE before transfer to PVDF. The blot was probed for Npr2 and FLAG and then stripped and reprobed for HA. B, Npr2-7A has a dominant-negative effect on Npr2. The . The 293T cells were transfected with 1 μg GFP plasmid, 2 μg FLAG-mouse-Npr2-WT, 1 μg FLAG-mouse-Npr2, and 1 μg HA-human-Npr2-7A or 2 μg HA-human-Npr2-7A. After 48 h, crude membranes were prepared and assayed for guanylyl cyclase activity. Values represent mean ± SEM from three experiments assayed in duplicate for n = 6.