Fig. 3.

KD of DDX3 reduces AKT activity, AKT-mediated GSK3β phosphorylation and ectopically expressed Snai1. (A,D,E,G) HEK293T cells were transfected with the indicated siRNA (200 nM) for 48 h (A,D,G), or treated with DMSO or 10 µM RK-33 for 24 h (E). In G, a plasmid encoding HA-tagged Snai1 (1 µg per well for six-well plates) was also transfected. (B,F) Embryos were injected at the one-cell stage with the indicated MO (12 ng each; 50 pg mRNA encoding HA-tagged Snai1 was co-injected in F), and cultured to stage ∼12.5. Cell or embryo lysates were processed for western blot analyses using the indicated antibodies. (C) Summary of the results of three independent experiments shown in B. Density of the target protein was normalized against that of β-actin, and fold change (DDX3 MO versus control MO) is shown. *P<0.05; **P<0.01 (unpaired t-tests). Data are mean±s.e.m.