Figure 2

Analysis of SV40 TAg expression and function in MOVCAR cells. A) Whole cell lysates of spontaneously transformed MOSEC lines (IF-5, ID-8 and IG-10) and MOVCAR cell lines (12-3, 5025, 5183, 5438, 5447, 5612 and 5009) were evaluated by immunoblot analysis to determine relative levels of SV40 TAg, p53 and Actin (loading control) protein expression. Lysates were also immunoprecipitated with anti-TAg antibody Pab 101 followed by immunoblot analysis of TAg and p53 protein present in the immunoprecipitates. B) Induction of p53 protein was evaluated by immunoblot following treatment of ID-8 and MOVCAR 5025, 5447 and 5612 cells with 200 nM etoposide for 0, 8 and 24 hr. C) Levels of Mdm2 gene expression in ID-8 and MOVCAR 5025, 5447 and 5612 cells following treatment with 200 nM etoposide for 0, 8 and 24 hr were evaluated by qRT-PCR. D) Cell cycle analysis was performed on ID-8 and MOVCAR 5025, 5447 and 5612 cells following treatment with 200 nM etoposide.