Transdux virus transduction reagent

The miR-195 coding sequence was cloned into the pMIRNA1 lentivectors (Human pre-microRNA Expression Construct Lenti-miR-195 MI0000489, Cat No: PMIRH195PA-1, SBI, USA) for expressing the miR-195 precursor (miR-195). Precursor Scramble control hairpin in pCDH-CMV-Scramble hairpin-EF1-copGFP (CD511B-1) was purchased from the same vendor (Cat No: PMIRH000PA-1, SBI, USA) for negative controls (miR-NC). The miRZip anti-microRNA Constructs (miRZip-195 anti-miR-195 microRNA construct, Cat No: MZIP195-PA-1) and miRZip control vectors (pGreenPuro Scramble Hairpin Control-Construct, Cat No: MZIP000-PA-1) were designed and cloned by SBI (USA). To package the construct, 293TN cells were transfected with miR-195/miR-NC or anti-195/anti-NC by pPACKH1 Packaging Plasmid Mix (Cat No: LV500A-1, SBI, USA), and then after 3 days, the virus particles were collected according to the packaging protocol of SBI with the Lenti-Concentin Virus Precipitation Solution (Cat No: LV810A-1, SBI, USA). DU145 and LNCaP cells were infected with TransDux virus transduction reagent (Cat No: LV850A-1, SBI, USA). The infected cells were isolated with a flow cytometer and cultured in 96-well plates.