Figure 3. Isolation of prostate CTCs from metastatic PCa patients using anti-CD45 immunomagnetic depletion.

2.5 ml blood from three metastatic PCa patients (> 50 CTCs/ 2.5 ml blood) was processed via ficoll density centrifugation and the PBMC fraction was collected. Immunomagnetic anti-CD45 depletion was performed on the obtained PBMCs and the remaining cells were washed, cytospunned onto the slides. Slides were stained for PSMA, EpCAM, sLe^x, and CXCR4 using the protocol as described in Figure S1. MDA, PC3, and KG1 cells were simultaneously stained as a control for the following markers: PSMA= Magenta, EpCAM= Yellow, HECA-452= Green, CXCR4= Red. All prostate CTCs expressed CXCR4, while, sLe^x expression was variable. The analysis of sLe^x intensity is shown in Figure 4.