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Fig. 6. Symplekin depletion inhibits ZONAB function. (A) A control HT29-16E clone and two clones expressing symplekin shRNA were grown without (white bars) or with tetracycline (grey bars) and then transfected with the ZONAB-specific luciferase reporter plasmids. Note the stimulation of the promoter upon symplekin depletion, indicating inhibition of the transcriptional activity of ZONAB. Values represent the mean ± s.e.m. of triplicates from three experiments (*P<0.05, Student's t-test compared with untreated clones). (B) Symplekin RiT-A5 clone and a control clone were grown in the absence or presence of the indicated concentrations of tetracycline for the indicated duration. Cells were transfected with equal amounts of reporter plasmids. 24 hours later, cells were lysed and luciferases were assayed. Ratios of the two luciferase activities were then calculated and compared between the different samples. Values are the mean ± s.e.m. of a representative experiment performed in quadruplicate. The results obtained with the symplekin RiT-A5 clone are expressed relative to the activity in the respective (untreated or tetracycline-treated) control sample.
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