Skip to main content
. 2014 Sep 22;9(2):355–364. doi: 10.1016/j.molonc.2014.09.005

Figure 4.

Figure 4

Essential roles of ZEB1 in RARA‐induced invasive phenotype. (A, B) Assessment of knockdown effects of shRNAs targeting ZEB1 and ZEB2. MCF10A/Tet‐on/TRE‐RARA cells infected with indicated shRNA viral vectors were cultured in the absence or presence of Dox for 3 days. ZEB1 and ZEB2 mRNA was analyzed by real‐time PCR analysis. The data were normalized by the amount of 18S rRNA and shown as a relative value compared with the control shRNA‐infected cells without Dox. Representative results of one of two independent experiments are shown. (C) Inhibition of RARA‐induced typical EMT marker transition by ZEB1 knockdown. Protein extracts from the ZEB1‐ or ZEB2‐knockdown cells were analyzed by immunoblotting. These cells were prepared as shown in (A) and (B), and cultured in Dox for 4 days with one passage. Tubulin was used as a loading control. (D) Inhibition of an RARA‐induced protrusive phenotype in Matrigel and collagen I cultures by ZEB1 knockdown. The ZEB1‐knockdown cells were prepared as shown in (A), and cultured in Dox for 2 days in 2D cultures and for 4 additional days in 3D cultures. Invasive protrusions of these cells in Matrigel and collagen I cultures were observed on day 4.