Figure 1: MCH-mediated ERK activation efficiently desensitizes. BHK-570 cells were transfected with plasmid DNA encoding MCHR1 an ERK activation assay performed on cells treated with MCH for up to 30 min. A second set of dishes was treated for MCH for 15 min and then subjected to a 30 min washout prior to the MCH time course (2nd exposure). (a) Cells were harvested in 2X Laemmli sample buffer, run on SDS-PAGE and western blots were performed with primary antibodies to phosphorylated and total ERK. (b) Densitometry was performed using NIH Image J Software where ERK activation was normalized against total ERK. Experiment shown is representative of three total experiments.