(a and b) Association of β-arrestin
with src and NK1R. KNRK-NK1R and KNRK-NK1Rδ325 cells were treated
with or without 10 nM SP for 5 min, crosslinked with DSP, and
immunoprecipitated with β-arrestin (β-arr) antibody, followed by
Western blotting with src (a) or NK1R (b)
antibodies. (c and d) Localization of
β-arrestin, src, and NK1R. KNRK-NK1R + ARR cells were incubated with
100 nM Alexa-SP (c) or 10 nM SP (d) for
60 min at 4°C, washed, and incubated at 37°C for 0–30 min. NK1R
was localized by Alexa-SP, β-arrestin with GFP, and src by
immunofluorescence with a Texas-red-conjugated secondary antibody.
Colocalization is shown in yellow in the merged images. Arrowheads
indicate plasma membrane, and arrows indicate intracellular
localization. (Scale bar = 10 μm.)