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Subsequently cells were rinsed twice with PBS, incubated in Alexa Fluor 488 goat anti-mouse following the manufacturer’s protocol (Invitrogen) for 2 hours at room temperature, rinsed again with PBS, and mounted with ProLong Gold Antifade Reagent (Invitrogen)

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in the case of PKA-mediated phosphorylation and desensitization, low nanomolar concentrations of agonist are sufficient to phosphorylate and desensitize the β2-AR, due to the amplification of the response through cyclic AMP generation and PKA activation