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Fig. 3. MAK is an activator of JNK. JNK activity was analyzed in embryonic
lysates by in vitro phosphorylation of GST-Jun(1-135). Four-cell embryos were
injected marginally four times with GFP, MAK or MAK-KD RNA (1.5 ng per
injection), or
N-Fz8 RNA (0.5 ng per injection) as a positive control.
Lysates were collected at stage 14 for JNK activity determination by western
analysis with anti-phospho-Jun antibodies. Assays were carried out in
duplicates, with 10 embryos in each experimental group. Anti-GST and
anti-ß-tubulin antibodies indicate loading.