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Figure 4


Fig. 4. Downregulation of symplekin in HT29-16E cells. (A) Regulated depletion of symplekin in the intestinal adenocarcinoma cell line HT29-16E. Clones derived from transfections with either control plasmid or the symplekin RNAi plasmid were analysed after 4 days of culture in tetracycline. The cells were lysed and the expression of the indicated proteins was analysed by immunoblotting. Actin was blotted as a loading control. Note that partial depletion of symplekin led to reduced expression of ZONAB-B. (B) Expression of ZONAB mRNA in HT29-16E cells. Control and symplekin RNAi HT29-16E cells were grown for 4 days in the presence or absence of tetracycline. Total RNA was purified and equal amounts were used for the synthesis of the first strand cDNA according to the manufacturer's instructions (Invitrogen). The level of ZONAB-B mRNA was analysed using semi-quantitative RT-PCR.





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