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Fig. 2. Anti-Rsk antibody 2 neutralizes Rsk activity in vitro and in vivo.
(A) The anti-Rsk serum 2 inhibits Rsk activity in vitro. Extracts were
prepared from immature oocytes (0) and unfertilized mature eggs (150).
Immunoprecipitates with anti-Rsk serum 1 and 2 from these extracts were
immunoblotted with anti-Rsk serum 2 (upper), or assayed for phosphorylation of
S6 peptide (lower). (B) The purified anti-Rsk antibody 2 inhibits Rsk
activity in vivo. Immature oocytes were injected with 3 ng of the anti-Rsk
antibody purified from the serum 2 (lane 6) or control IgG from preimmune
serum 2 (lane 5), or not injected (lane 1-4); oocytes were then treated with
1-MeAde. Extracts were prepared at the indicated times after 1-MeAde addition,
and immunoblotted with anti-MAPK antibody (third panel) and anti-active MAPK
(fourth panel). Immunoprecipitates with the anti-Rsk serum 1 were
immunoblotted with anti-Rsk serum 2 (second panel), and assayed for
phosphorylation of GST-S6 (first panel). In lane 4 (210F), mature eggs were
inseminated 150 minutes after 1-MeAde addition. In lane 6, Rsk was present in
the U form but its activity was almost undetectable.
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