Identification and Characterization of ZEL-H16 as a Novel Agonist of the Histamine H3 Receptor
Figure 6
Localization of internalized H3R-EGFP stably expressed in HEK-293 cells and recycling of internalized H3R to the cell surface.
A. HEK-293 cells stably expressing H3R-EGFP were incubated with or without 20 µM ZEL-H16 and 500 µM histamine in the presence of 100 g/ml Alexa Fluor546-labeled transferrin for 45 min at 37°C. B. H3R-EGFP expressing cells were treated with 100 µg/ml cycloheximide and 20 µM ZEL-H16 or 500 µM histamine at 37°C for 30 min, followed by the removal of residual agonists by washing, and further incubation in the presence of cycloheximide for the indicated time periods. The internalized receptors were recycled to the plasma membrane within 1 h after histamine removal and 3 h after ZEL-H16 removal. All pictures shown are representative of at least three independent experiments.