Vybrant did cell labeling dye

Early study has shown that DAB2IP gene knockdown in PC3 cell, a poorly-metastatic line, increases its metastatic potential (also called as DAB2IP-knockdown PC3 cells or KD cells)²⁴ . These stable cell lines expressing dual reporter genes (GFP and luciferase) were maintained in RPMI1640 medium (Invitrogen, Carlsbad, CA, USA) containing 5% PBS as previous described²⁴ . Migration assays were performed in Transwell dishes (Corning Costar, Cambridge, MA, USA) as described earlier^{73 (link)}. For tracking the cell migration in vivo, both PC3 and KD were labeled with Qtracker® labeling kit (Life technologies, Carlsbad, CA, USA) by following manufacture's instruction as illustrated earlier^{74 (link)}. For histological study, Vybrant DiD cell labeling dye (Thermo Fisher Scientific, Waltham, MA, USA) was used to track cancer cells according to the product information for observation under microscope^{75 (link)}.

Various antibodies were purchased from the following suppliers: HIV-1 Tat (#ab43014) from Abcam, Cambridge, MA, USA; against phospho-DRP1 (#3455), Alix (#2171), GRP94 (#2104), and p-eNOS (#9571) from Cell Signaling Technology, Danvers MA, USA; against CD81 (#sc-23962), CD63 (#sc-5275), and TSG101 (#sc-7964) from Santa Cruz Biotechnology, Dallas, TX, USA; β-actin (#A5441) from Sigma-Aldrich; total DRP-1 (#611112), total eNOS (#610296) from BD Transduction Laboratory, San Jose, CA, USA. Vybrant DiD cell-labeling dye for exosome staining was purchased from Molecular Probes (Eugene, OR, USA). Dihydroethidium (DHE) (Thermo Fisher Scientific, Waltham, MA, USA) was used to detect cytosolic superoxide and MitoSOX Red (Molecular Probes) for the detection of mitochondrial superoxide. To label mitochondria, MitoTracker Red and MitoTracker Green were used (Molecular Probes). Phorbol 12-myristate 13-acetate (PMA), PX866, Dynasore, and Cytochalasin D were from Cayman Chemical (Ann Arbor, MI), Ionomycin was from Sigma-Aldrich, and recombinant human IL-2 (rIL-2) was from Miltenyi Biotec (Auburn, CA, USA). GW4896 was from Selleckchem (Houston, TX).