1. A B C D eIF3e shRNA 205 clones Control shRNA 3 7 eIF3e b-actin
eIF3e
b-actin C D
eIF3e shRNA
clone 3
eIF3e shRNA
clone 7
Control shRNA
Day 4
Day 10
Reduced eIF3e expression causes epithelial-to-mesenchymal transition in MCF-12A cells. A) Confirmation of eIF3e knockdown in individual
clones of stably-transduced MCF-12A cells; eIF3e and b-actin levels were determined by Western-blot analyses. B) Total mRNA was harvested
from MCF-12A cells that harbor a non-silencing control shRNA or the eIF3e shRNA and was subsequently used as templates quantitative RT-PCR
for the epithelial marker E-cadherin, the mesenchymal marker N-cadherin, and the EMT regulator Snail1, as well as eIF3e and GAPDH; summary
of three independent experiments. C) Reduced eIF3e expression enhances migration of MCF-12A breast epithelial cells. The migration of
MCF-12A cells harboring a non-silencing control shRNA or eIF3e shRNA toward serum were determined by transwell migration assays; graph is a
summary of two independent experiments. D) Reduced expression of eIF3e disrupts mammary acinar structure. MCF-12A cells harboring a
non-silencing control shRNA or eIF3e shRNA were grown on Matrigel and monitored for development of three-dimensional structures;
photographs taken on the days indicated.