Figure 2.

PC-3M-Pro4 with enhanced epithelial–mesenchymal transition (EMT) and cancer stem-like cell (CSC) traits in comparison with PC-3 have stronger extravasation and metastatic tumor-initiating capacities. (a) PC-3-mCherry and PC-3M-Pro4-mCherry were injected into ZF vasculature. Confocal images were acquired at the metastatic site at 1, 2 and 4 dpi. Green, vessels. Red, cancer cells. Scale bar=50um. (b–c) Extravasation and cancer cell burden at the metastatic site were analyzed. For extravasation analysis, % of ZF with more than 1 cell extravasated from the caudal vein and invaded into neighboring tissue was counted. For cancer cell burden quantification, total fluorescence of mCherry was measured using a ZF-4 pixel counting software. Experiment was independently repeated 2 times with 30 fish per group. Error bar presented as mean ±SEM. (d–e) The expression of EMT markers and stemness markers in PC-3 and PC-3M-Pro4 in culture was measured by qPCR. Experiment was independently repeated 3 times. Data were presented as mean ± SD.