Figure 5. FNIP1 interacts with ER Ca 2... | Rockefeller University Press

Figure 5.

FNIP1 interacts with ER Ca2+-ATPase SERCA. (A and B) Co-IP experiments were performed by co-transfecting HA-SERCA2b and Flag-FNIP1 in HEK293T cells, as indicated at the top. Extracts (input) from the HEK293T cells and proteins from the IP were analyzed by immunoblotting (IB). Representative results for co-IP are shown. n = 3 independent experiments. (C) GFP-FNIP1 and Mcherry-SERCA2b colocalized in the cytoplasm. Scale bar: 100 μm. (D) Top: Schematic diagrams of different HA-tagged SERCA2b constructs. Bottom: Schematic diagrams of different Flag-tagged FNIP1 constructs. a.a., amino acid. (E) Deletion of the P or NB region abolishes the coimmunoprecipitation of FNIP1 and SERCA2b. Representative results for co-IP are shown. n = 3 independent experiments. (F) Deletion of the A region abolishes the co-IP of SERCA2b and FNIP1. Representative results for co-IP are shown. n = 3 independent experiments. (G) The schematic depicts a proposed model for the FNIP1-SERCA2b interaction. (H) Co-IP of endogenous FNIP1 and SERCA2 using iWAT from male WT mice treated with NaCl or CL316,243 (1 mg/kg) for 1 h. Representative results for co-IP are shown. n = 3 independent experiments. Data are representative of three independent experiments (A–C, E, F, and H). Source data are available for this figure: SourceData F5.

FNIP1 interacts with ER Ca ²⁺ -ATPase SERCA. (A and B) Co-IP experiments were performed by co-transfecting HA-SERCA2b and Flag-FNIP1 in HEK293T cells, as indicated at the top. Extracts (input) from the HEK293T cells and proteins from the IP were analyzed by immunoblotting (IB). Representative results for co-IP are shown. n = 3 independent experiments. (C) GFP-FNIP1 and Mcherry-SERCA2b colocalized in the cytoplasm. Scale bar: 100 μm. (D) Top: Schematic diagrams of different HA-tagged SERCA2b constructs. Bottom: Schematic diagrams of different Flag-tagged FNIP1 constructs. a.a., amino acid. (E) Deletion of the P or NB region abolishes the coimmunoprecipitation of FNIP1 and SERCA2b. Representative results for co-IP are shown. n = 3 independent experiments. (F) Deletion of the A region abolishes the co-IP of SERCA2b and FNIP1. Representative results for co-IP are shown. n = 3 independent experiments. (G) The schematic depicts a proposed model for the FNIP1-SERCA2b interaction. (H) Co-IP of endogenous FNIP1 and SERCA2 using iWAT from male WT mice treated with NaCl or CL316,243 (1 mg/kg) for 1 h. Representative results for co-IP are shown. n = 3 independent experiments. Data are representative of three independent experiments (A–C, E, F, and H). Source data are available for this figure: SourceData F5.