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. 2001 Nov;21(21):7429–7441. doi: 10.1128/MCB.21.21.7429-7441.2001

FIG. 1.

FIG. 1

FIG. 1

Ephrin-B1 stimulation of EphB2 leads to the down regulation of the ERK1/2 MAPK signaling pathway. (A and B) Serum-starved (right panels) or growing (10% FBS) (left panels) parental NG108 or NG-EphB2 cells were stimulated with 2 μg of clustered Fc-ephrin-B1 per ml for the indicated time points and lysed directly in 2x SDS-PAGE sample buffer. The lysates were electrophoresed and blotted (WB) with antibodies against phosphorylated ERK1/2 (A, top panels), or phosphorylated MEK1 (B, top panels). The blots were stripped and reprobed for total ERK1 or MEK1 (bottom panels, A and B, respectively). (C) Graphical representation of phospho-ERK1/2 and phospho -MEK1 from panels A and B relative to total levels. (D) Time course of EphB2 tyrosine phosphorylation in NG108 cells. NG108 cells were serum starved (lanes −serum) or grown in the presence of serum (lanes +serum) and stimulated with 2 μg of clustered Fc-ephrin-B1 per ml as indicated. Immunoprecipitated (IP) EphB2 was then resolved by SDS-PAGE and probed with anti-pTyr antibodies (upper panel). Blots were subsequently stripped and reprobed with crude anti-EphB2 sera (lower panel).