FIG 6.

Both hAgo2 and AUF1(s) are required to downregulate DICER1 expression. (A) Western blotting of DICER1 protein using samples from HeLa cells stably transfected with shRNA targeting AUF1 (two different shRNA clones, I and II) and transiently transfected with an expression plasmid for hAgo2. (B) Luciferase activity was measured using samples from HeLa cells with pIS1-DICER1 long AREM. (C) Coimmunoprecipitation of hAgo2 and AUF1 proteins. Forty-eight hours after transfection of HEK293T cells with different plasmids, as indicated, cell lysates from these cells were directly analyzed by Western blotting (input) for expression control or immunoprecipitated with the anti-Myc antibody prior to Western blotting (bottom). AUF1 p45 protein was marked by an arrow. Results in panel B are the means (±SD) from three independent experiments.