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Figure 8


Fig. 8. Caveolin-1 downregulated survivin expression in cells with enhanced or constitutive activity via the Wnt pathway. (A) HEK293T cells were transfected with either pLacIOP (-) or pLacIOP-caveolin-1 (+) and then cultured either in normal medium (control) or conditioned medium containing Wnt3a for 24 hours (see Materials and Methods for details). Survivin, caveolin-1 and actin protein levels were assessed by western blotting. (B) Clones of DLD1 colon adenocarcinoma cells stably transfected with either pLacIOP (M1) or pLacIOP-caveolin-1 (C4) were described previously (Bender et al., 2000). Cell extracts were prepared post-induction with 1 mM IPTG for 24 hours. Survivin, caveolin-1 and actin expression were assessed by western blotting. Residual survivin level was 0.39±0.10 (mean ± s.e.m.; *P<0.05), as determined in three independent experiments. (C) Survivin mRNA content in DLD1 clones was analyzed by RT-PCR. Residual survivin mRNA level was 0.49±0.18 (mean ± s.e.m.; #P<0.05), as determined in three independent experiments.





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