(A). Knockdown of SNAI1, ZEB1, TWIST2, or a triple SZT knockdown in PC-3/S cells was associated with fewer cells with fibroblastoid morphologies and a gain in the expression of E-cadherin, most evident in ZEB1 knockdowns (single or triple SZT). Scale bars: 20 μm. (B) Knockdown in mesenchymal-like PC-3/S cells of SNAI1, ZEB1, or a triple SZT knockdown caused an upregulation of E-cadherin, as determined by Western blotting, with the strongest effect observed in the triple knockdown. (C) Knockdown of SNAI1, ZEB1, TWIST2, or a triple SZT knockdown caused a diminished invasive capacity of PC-3/S cells in Transwell-Matrigel assays, with the triple SZT knockdown showing the strongest effects. (D) Knockdown of SNAI1, ZEB1, TWIST2, or a triple SZT knockdown caused a gain in the capacity of PC-3/S cells to grow spheroids, with the triple knockdown showing the strongest effects. (E) Knockdown of SNAI1, ZEB1, TWIST2, or a triple SZT knockdown in mesenchymal-like PC-3/S cells caused an upregulation of the epithelial genes CDH1, EPCAM, and DSP and of the self-renewal/ pluripotency genes LIN28, SOX2, MYC, and KLF4, most evident for the triple SZT knockdown. Real-time RT-PCR values, determined by the ΔΔCp method, are represented as a heat map with pseudocoloring ranging from green (underexpressed relative to values in control PC-3/S cell) to red (overexpressed relative to control PC- 3/S cells). Controls were puromycin-selected PC-3/S cells bearing control pLK0-scrambled lentiviral vector. Results are expressed as mean ± SEM. *P < 0.05; **P < 0.01.