FIGURE 3.

Generation and characterization of tagged recombinant human RLN2 peptide. A, schematic design of recombinant RLN2 peptides with an N-terminal Myc tag and a His₆ tag at four different positions of the mature peptides (RLN2 His1-4). The mature B and A chains are indicated by a rectangle box, whereas the C domain is indicated by a line. Myc and His₆ epitopes are indicated by a light blue rectangle and a yellow rectangle, respectively. B, Western blotting analysis of tagged RLN2 peptides in conditioned media of transfected HEK293T cells using an anti-relaxin antibody (Calbiochem; upper panel) or an anti-Myc antibody (lower panel) under reducing conditions. The molecular weight standard is shown on the left. C, stimulation of cAMP production by RLN2 His1 and RLN2 His2 peptides in LGR7-expressing HEK293T cells.