FIG 5.

Downregulation of DICER1 by AUF1(s) requires hAgo2. (A) Western blot of various proteins involved in the miRNA pathway using samples from HeLa cells stably transfected with shRNA targeting AUF1 (two different shRNA clones, I and II). (B) Western blotting of DICER1 protein using samples from HeLa cells transiently transfected with the expression plasmids for one of the four different isoforms of AUF1 (p37, p40, p42, or p45). The expected proteins were marked by arrows, while the smaller proteins were probably the degraded products. (C) Western blot of DICER1 and hAgo2 proteins using samples from HeLa cells transiently transfected with different amounts of either vector or the plasmids expressing four different forms of AUF1, as indicated. (D) Western blot of various proteins involved in the miRNA pathway using samples from HeLa cells transiently transfected with the expression plasmids for the four isoforms of AUF1. (E) Western blot of DICER1 protein using samples from A549 cells stably transfected with sgRNA targeting hAgo2 (two different clones [2-19 and 1-14] from A549 cells) and transiently transfected with expression plasmids for four different forms of AUF1. (F) Western blotting of DICER1 protein using samples from HeLa cells stably transfected with shRNA targeting hAgo2 (two different shRNA clones, +6 and +8) and transiently transfected with expression plasmids for four different forms of AUF1.