Figure 5. Low p27Kip1 expression in T24T cells mediated JNK/c-Jun phosphorylation, EGFR up-regulation and increased anchorage-independent growth.
(A) Total cell lysates from T24 and T24T bladder cancer cells were subjected to Western blotting as indicated. β-Actin was used as a protein loading control. (B and C) Anchorage-independent growth of T24 and T24T cells was determined by soft agar assay. Colonies were photographed and counted, and values are means±S.D. for triplicate assays. (D) T24T cells were pretreated with or without SP600125, and then extracted for determination of protein expression by Western blotting. (E and F) T24T cells were pretreated with 25 μM SP600125 for 4 h and then subjected to anchorage-independent growth. Colonies were photographed and counted. The asterisk (*) indicates a significant decrease in anchorage-independent growth of T24T pretreated with SP600125 compared with T24T cells pretreated with 0.1% DMSO control (P<0.01).