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Fig. 6. Mitogenic properties of mAb16-39 are dependent on ERK activity. (A) Effect of mAb16-39 on ERK activation. 2-C4 cells were serum starved for 24 hours and treated with 4 nM normal rat IgG (–) or mAb16-39 (+) for 5 minutes or 3 hours. Equal amounts of total cell lysate were probed with phospho-specific anti-ERK1/2 antibody (ERK1/2-P). The blots were stripped and reprobed with anti-ERK antibody (ERK2). (B) Quantification of data presented in A. The intensity of the bands in the anti-phospho-ERK1/2 blot was normalized against the amounts of MAPK and expressed as fold activation over basal levels (IgG-treated). (C) Effect of the MEK inhibitor PD98059 on the mitogenic activity induced by mAb16-39. Serum-deprived 2-C4 cells were left untreated or were treated with 4 nM IgG or mAb16-39 for 2 days in the presence or absence of PD98059 (50 µM) or wortmannin (100 nM). [³H]thymidine uptake assay was performed as in Fig. 5, and the data are presented as the mean±s.e.m.

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