Figure 5

TDAG51 expression is induced by ERK pathway activation. HME16C cell lines infected with pLRT empty vector (pLRT), Ras^V12 (V12), or Ras^V12S35 (S35) were treated with 0.2% DMSO vehicle or 20 μM PD98059 for 24 hours, followed by western blotting for TDAG51. The blot was stripped and probed with an alpha tubulin monoclonal antibody (Alpha tubulin) to demonstrate equal protein loading. Band intensities were determined using Image J software (NIH, Bethesda, MD) and TDAG51 values were normalized using alpha tubulin values. Fold expression of PD98059-treated relative to DMSO-treated contols: pLRT (0.6); V12 (0.6); S35 (0.3). Similar experiments to that shown was performed 1 additional time with similar results.