Figure 6
From: ZEB1 expression in type I vs type II endometrial cancers: a marker of aggressive disease
Partial knockdown of ZEB1 in Hec50co cells does not restore E-cadherin, but significantly reduces migratory behavior. Three shRNAs designed to knock down ZEB1 and a nonsilencing control were tested using the RNA Intro GIPZ shRNAmir kit (Open Biosystems). (a) Real-time RT-PCR of ZEB1 RNA levels is shown. G10 shRNAmir was most efficient. (b) Short hairpin RNA G10 and a nonsilencing control were each stably transfected into Hec50co cells and whole cell extract from the respective pooled populations and wild-type Hec50co cells were prepared. An immunoblot of 150 μg of protein from each cell line was probed with antibodies specific for ZEB1, E-cadherin, and α-tubulin. (c) In Boyden chamber migration assays Hec50co-ZEB1shRNA G10 cells were significantly less migratory than Hec50co cells stably expressing nonsilencing control or wild-type Hec50co cells as determined by a one-way ANOVA with Tukey's multiple comparison test (P=0.152).
