Figure 4. Nesfatin-1 specifically binds to the cell surface of NB41A3 cells and mouse hypothalamus.

NB41A3 cells were plated in a 24-well format, and the total bound activity of ¹²⁵I labeled Nesfatin-1 on collected NB41A3 cells was counted (A). The total bound activity of ¹²⁵I labeled Nesfatin-1 on the membrane fraction of mouse hypothalamus was counted (B). Non specific binding was observed in the presence of excess of unlabeled Nesfatin-1. Saturation binding isotherm with Hill slope for specific binding after excluding nonspecific binding is shown. The Kd value and Bmax values were calculated by curve fitting program of Prism 5. Scatchard plot analysis is shown in the right panel.