Figure 3

Inhibition of AKT activation by ODAM expression in human melanoma cell lines. A,B) Western blot analysis of AKT activation in total cell lysates from control and ODAM-expressing A375 and C8161 melanoma cells grown under normal culture conditions. Multiple blots from the same lysate sets were probed sequentially with the indicated antibodies. ODAM expression was detected by immunoprecipitation from cell culture supernatants C) Quantitative real time RT-PCR analysis of PTEN mRNA levels in control and ODAM-expressing cells growing under normal culture conditions. Values for ODAM-expressing cells represent the mean ± 1 S.D. from five independent determinations expressed relative to values from control cells assayed concurrently. D) Analysis of PTEN protein synthesis in control and ODAM-expressing A375 cells by metabolic labeling and immunoprecipitation as given in the methods.