Figure 4.
Figure 4. hCLR(V205del) does not interact with RAMPs. (A) Representative saturation isotherms from BRET assays of transiently transfected HEK293T cells with hCLR (top graph) or hCLR(V205) (bottom graph; fused to renilla luciferase) and increasing amounts of hRAMP2 (fused to eYFP). Experiments were performed at least three times and a representative experiment is shown. (B) PLA in transiently transfected HEK293T cells. Red indicates positive PLA signal, blue is nuclear DAPI stain, green is plasma membrane β-catenin. Negative controls included no transfection of hRAMP2. Graph depicts quantitation of signal, with asterisks denoting significant differences compared with control; ***, P < 0.001. n = 2 independent experiments, in duplicate, with 20 cells quantitated per condition. Bar, 10 µm.

hCLR(V205del) does not interact with RAMPs. (A) Representative saturation isotherms from BRET assays of transiently transfected HEK293T cells with hCLR (top graph) or hCLR(V205) (bottom graph; fused to renilla luciferase) and increasing amounts of hRAMP2 (fused to eYFP). Experiments were performed at least three times and a representative experiment is shown. (B) PLA in transiently transfected HEK293T cells. Red indicates positive PLA signal, blue is nuclear DAPI stain, green is plasma membrane β-catenin. Negative controls included no transfection of hRAMP2. Graph depicts quantitation of signal, with asterisks denoting significant differences compared with control; ***, P < 0.001. n = 2 independent experiments, in duplicate, with 20 cells quantitated per condition. Bar, 10 µm.

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