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Figure 1.
FIG. 1. Dynamic nature of endogenous DLK, JIP1, and JNK
interaction in stimulated HN33 cells. A, extracts were prepared
from HN33 cells (1 x 10^7) treated with or without 400 nM
okadaic acid (Ok. Acid) for 3 h. JIP1 complexes were
immunoprecipitated from the cell lysates using JIP1-specific
antibody, separated by SDS-PAGE, and analyzed by immunoblotting
with anti-DLK and anti-JNK antibody. Immunoprecipitation without
JIP1 antibody was used as control. Corresponding lysates were
immunoblotted as indicated to determine the endogenous
expression of DLK, JIP1, and JNK. B, HN33 cells were transfected
with FLAG-JIP1. Cells were treated with either kainic acid or
okadaic acid as indicated. JIP1 was immunoprecipitated using
JIP1 antibody and immune complexes were analyzed for the
presence of DLK. C, HN33 cells were transfected with FLAG-JIP1,
metabolically labeled with orthophosphate, and then treated with
or without kainic acid for 60 min. JIP1 was immunoprecipitated
using JIP1 antibody and immune complexes were analyzed after
SDS-PAGE by autoradiography for incorporation of phosphate into
JIP1 or by immunoblotting for the presence of DLK or JNK. All
experiments were repeated 3 times with similar results.
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