FIGURE 5.

Dominant-negative deadenylases alleviate PUM repression. A, expression of a dominant-negative CNOT8 mt inhibits PUM repression in HEK293 cells. Graph of percent repression, relative to the RnLUC, of the indicated reporters in cells transfected with increasing amounts of plasmid expressing CNOT8 mt protein is shown. B, expression of the Halotag-CNOT8 mt fusion protein was confirmed by fluorescent TMR labeling and detection on a SDS-PAGE gel from the samples in panel A. Halotag alone was used to balance the transfected mass of DNA and is therefore also detected. C, dominant-negative CNOT7 mt inhibits PUM repression. Graph of percent repression relative to RnLUC control of the indicated reporters in cells transfected with increasing amounts of plasmid expressing CNOT7 mt protein. D, expression of Halotag-CNOT7 mt fusions was confirmed by TMR detection on SDS-PAGE from the samples in panel C. Halotag alone was used to balance the transfected mass of DNA and is therefore also detected. In all graphs, mean values are graphed with mean ± S.E.