Analyzing the Role of Receptor Internalization in the Regulation of Melanin-Concentrating Hormone Signaling

Figure 4

Overexpression of 𝛽-arrestins differentially influences MCHR1 internalization. BHK-570 cells were transfected with VSVg-MCHR1 and either 𝛽 arrestin-1 or 𝛽 arrestin-2. (a) Cells were treated with 1 𝜇M MCH or vehicle for 30 min and internalization was measured utilizing a cell-based ELISA. Data from Figure 3 was replotted. Ave ± SEM plotted. ^∗∗𝑃<0.005, ^∗𝑃<0.01. VSVg-MCHR1 and GFP 𝛽-arrestin 1-expressing cells (b) and VSVg-MCHR1 and GFP 𝛽-arrestin 2-expressing cells (c) were treated with 1 𝜇M MCH or vehicle for 30 min, then fixed, and immunostained with anti-VSVg antibody. Fluorescence microscopy was performed and overlay images compared.

(a)

(b)

(c)