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Figure 5.
Arrestin-2 and -3 can homo- and heteroassociate in COS-1
cells. A, co-immunoprecipitation experiment showing wild type
arrestin-2 associating with itself. Mutation of the two
crystallographically defined IP[6]-binding sites on arrestin-2,
either independently or combined, disrupts self-association.
HA-tagged wild type and mutant arrestin-2 proteins were
overexpressed in COS-1 cells with or without FLAG-tagged wild
type arrestin-2. HA-arrestins were immunoprecipitated and
blotted for FLAG-arrestin-2. Immunoprecipitate (IP) and lysate
expression levels are shown. B and C, co-immunoprecipitation
experiments showing arrestin-2 mutants effectively interacting
with known binding partners. HA-tagged wild type and mutant
arrestin-2 proteins were overexpressed in COS-1 cells.
HA-arrestins were immunoprecipitated and blotted for clathrin,
β[2]-adaptin, and ERK2. Immunoprecipitate and lysate expression
levels are shown. D, co-immunoprecipitation experiment showing
that wild type arrestin-2 and -3 can homo- and heteroassociate.
HA-tagged arrestin-2 and -3 proteins were overexpressed in COS-1
cells with or without FLAG-tagged arrestin-2 or -3. HA-arrestins
were immunoprecipitated and blotted for FLAG-arrestins.
-indicates mock transfection. IB, immunoblot; WT, wild type;
arr2, arrestin-2; arr3, arrestin-3.
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