Detection of MAP and JNK phosphorylations
κ-Neuro-2A cells were cultured in 60 mm plates for 48 h in the presence
or absence of U50,488H (20 μΜ) for 15min and 6 h at 37oC. Cell monolayers were rinsed with PBS following the
procedure as described by (43). Where
necessary cells were exposed to the ERK1, 2 inhibitor PD98059 (20 μΜ for
45 min), or the JNK inhibitor SP600125 (20 μΜ for 30 min) prior to
agonist treatment. The proteins were resolved in 10 % SDS-PAGE and
visualized by immunoblotting with the appropriate antibodies as
described previously by (50).