ASK1, MKK4, and JNK3 interact with both domains of arrestin.
A-C, HEK-293A cells were transfected with HA-ASK-NLS or GFP-JNK3
individually or with FLAG-tagged full-length arrestin3 (FLAG-Arr3), its N-
(FLAG-Arr3-N) or C-domain (FLAG-Arr3-C), arrestin2-NES+ (Arr2-NES), arrestin2
N-domain-NES (Arr2-N-NES), or arrestin2 C-domain-NES (Arr2-C-NES).
Immunocytochemistry was done as described in
Fig. 2C. GFP-JNK3 was
visualized by its intrinsic fluorescence. Representative images are shown. The
localization of GFP-JNK and HA-ASK1-NLS was quantified, as described in the
legend to in Fig. 2C.
Means ± S.D. (n = 3) of the fraction of cells with more JNK3
or ASK1 in the cytoplasm than in the nucleus also shown. (****,
p < 0.0001). D-F, COS-7 cells were transfected with
HA-ASK1, HA-MKK4, or GFP-JNK3 alone, or together with FLAG-tagged arrestin3
and its domains. Immunoprecipitation and Western blot analysis were performed
as described in the legend to Fig.
2A.