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Figure 6 | BMC Cell Biology

Figure 6

From: Delayed internalization and lack of recycling in a beta2-adrenergic receptor fused to the G protein alpha-subunit

Figure 6

Dual-localization of co-expressed β 2 -adrenergic receptors after antagonist-mediated recycling. HEK293 cells stably expressing Flag-β2AR-Gαs receptor were transiently transfected with HA-β2AR construct. After 48 hours, receptor internalization was induced by incubating cells with 1 μM isoproterenol for 5 hours. 1 μM propranolol was added (Agonist + Antagonist) or not (Agonist) for a further 60 minutes to stop receptor endocytosis. Fixed and immuno-stained cells were subjected to epifluorescence microscopy, as described in Methods, to visualize the sub-cellular localization of HA-β2AR (Alexa-Fluor 594 staining; red) and Flag-β2AR-Gαs (Alexa-Fluor 488 staining; green). Scale bars, 10 μm. Note that the antagonist allows the membrane return of the wild type receptor but not of the chimeric protein. Photographs represent the typical situation observed in 4 independent experiments.

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