Figure 4

Expression of TDAG51 in HME16C cell lines in the absence or presence of the EGFR inibition. Western blotting of TDAG51 in HME16C cell lines. (A) Twenty-five micrograms of protein from HME16C cellular lysates were resolved by SDS-PAGE and probed with an anti-TDAG51 monoclonal antibody (TDAG51). Equal protein loading was verified by stripping the blot and probing with an alpha tubulin monoclonal antibody (Alpha tubulin). Band intensities were determined using Image J software (NIH, Bethesda, MD) and TDAG51 values were normalized using alpha tubulin values. Fold expression of each band relative to HME16C parental: HME16C parental, 1.0; pLRT, 1.0; V12, 3.0; G37, 2.3; S35, 2.4; C40, 1.3; Rlf-CAAX, 0.9. Similar experiments to that shown were performed 2 additional times with similar results. (B) HME16C cell lines were treated with DMSO vehicle or 0.25 μM PD153035 EGFR-specific inhibitor for 48 hours, and cell lysates were prepared. Twenty five micrograms of protein were resolved by SDS-PAGE and western blotted with anti-TDAG51 mAb (TDAG51). The blot was subsequently stripped and probed with an alpha tubulin mAb (Alpha tubulin). Band intensities were determined using Image J software (NIH, Bethesda, MD) and TDAG51 values were normalized using alpha tubulin values. DMSO-treated cells are indicated by a -, and PD1535035-treated cells are indicated by a +. Fold expression relative to DMSO-treated pLRT: pLRT -(1.0), +(0.2); V12 -(2.3), +(2.6); G37 -(0.8), +(0.3); S35 -(1.4), +(0.9); C40 -(1.1), +(0.6); Rlf-CAAX -(1.6), +(1.0). Similar experiments to that shown were performed 2 additional times with similar results.