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Western Blot Analysis {#sec3dot8-molecules-22-00702} -------------------------- The expression of uPAR was examined in isolated ASCs/PC3-NT and ASCs/PC3-Cyst. First, cells were cultured on a 12-well plate at 5 × 10^5^ cells/well for 3 days. Then, cells were lysed in M-PER buffer (Thermo Fisher Scientific, Waltham, MA, USA), containing protease and phosphatase inhibitors (Sigma-Aldrich). The protein concentration was determined with a DC protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA). Equal amounts of protein were separated by SDS-PAGE, and were transferred to a PVDF membrane (Immobilon-P; Millipore, Billerica, MA, USA) in a buffer containing 25 mM Tris, 190 mM glycine, and 20% methanol. The PVDF membrane was blocked for 1 h with Tris-buffered saline (TBS) containing 5% nonfat dry milk. After washing with TBS containing 0.1% Tween 20 (TBST), the membrane was incubated with primary antibody (1:1000 in TBST) for 1 h at room temperature. 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