Fig. 3. Phosphorylation of c-Jun by activated JNK is adhesion independent over a
range of anisomycin concentrations. Serum-starved detached NIH 3T3 cells were
either plated onto fibronectin (Fn)-coated dishes or maintained in suspension
(Sus) for 2 hours in DMEM/BSA. At this time point, cells were stimulated (A)
with increasing doses of anisomycin (0, 5, 20, 50 and 100 ng/ml) for 30
minutes or (B) with 50 ng/ml anisomycin for the times indicated. Under each
condition, cells were lysed in modified RIPA buffer and lysates analyzed by
western blotting for phosphoserine-63 c-Jun (Phospho63-cJun), phosphoserine-73
c-Jun (Phospho 73-cJun) and total c-Jun. The results shown are representative
of three independent experiments.
