Fig. 5. Anisomycin treatment activates p38 in an anchorage-independent manner. Serum-starved detached NIH 3T3 cells were either maintained in suspension (Sus) or plated onto fibronectin (Fn)-coated dishes for 2 hours in DMEM/BSA. (A) At the 2 hour time point, cells were stimulated appropriately with increasing doses of anisomycin (0, 25, 50 and 200 ng/ml) for the 30 minutes. (B) Cells were treated with 50 ng/ml anisomycin for the times indicated. Under each condition, cells were lysed in modified RIPA buffer and cell lysates probed with antibodies to phosphorylated and total p38.