Figure 3.

Knockdown of Zeb1 in PC-3 prostate cancer cells increases cell invasion. (A,B) Transwells were coated with 4 µg/well of Matrigel. Cells (5 × 10⁴/well) were added to wells and either control media (0% fetal bovine serum [FBS]) or chemoattractant media (5% FBS) was added and cells were allowed to invade for 24 h. Cells were fixed with 1% glutaraldehyde and mounted with DAPI-containing mounting media. (C,D) For spheroid invasion assays, cells were seeded onto ultra-low attachment plates and allowed to grow for 96 h to create spheroids. Matrigel was then added and invasion was quantified after 48 h. Representative images are shown for each assay; with invasion calculated based on 5 high-powered fields of view (HP-FOV) per well. Black scale bars = 100 µm, white scale bars = 300 µm. Data is presented as the mean ± standard error of the mean (SEM) (n = 3). α = significantly different than PC-3 control (ctrl) no doxycycline (Dox). β = significantly different than ctrl with Dox. δ = significantly different than Zeb1^KD (Zeb1 knockdown) no Dox (p ≤ 0.05).