Prl 1 plasmid

PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. PRL-1 lentiviral plasmid was purchased (SeouLin Bioscience, Seongnam, Korea). pLenti-RSV-EF1α vector including PRL-1 was constructed with a C-terminal GFP as well as the antibiotic puromycin. AMAXA pCMV-GFP vector was contained (Lonza, Basel, Switzerland). The resulting plasmid was confirmed by DNA sequencing. Naïve PD-MSCs (6 × 10 4 cells/cm 2 ) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by 2 µg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 in a humidified atmosphere containing 5% CO 2 .

The PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. The CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. The PRL-1 lentiviral plasmid was purchased from SeouLin Bioscience (Seongnam, Korea). The pLenti-RSV-EF1α vector including PRL-1 was constructed with a Cterminal GFP as well as the antibiotic puromycin. The AMAXA pCMV-GFP vector was obtained from Lonza (Basel, Switzerland). The resulting plasmid was con rmed by DNA sequencing. Naïve PD-MSCs (6x10 4 cells/cm 2 ) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by using 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by using 2 µg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 ℃ in a humidi ed atmosphere containing 5% CO 2 .