Cardiomyocytes (H9C2 cell line) were purchased from Cell Bank of Chinese Academy of Sciences. Cells were cultured in DMEM supplemented with 1.5 g/l NaHCO3, 10% FBS (Gibco, Invitrogen, U.S.A.), 1% glutamine, and 1% penicillin-streptomycin in 5% CO2 incubator under 37°C. H9C2 cells were treated with LPS for 24 h. MiR-25 mimic, miR-25 inhibitor, and pcDNA-PTEN were synthesized by GenePharma (Shanghai, China). Lipofectamine 2000 (Invitrogen) was used to perform cell transfection according to the manufacturer’s instructions.
Pcdna pten
Manufactured by GenePharma
PcDNA-PTEN is a plasmid vector used in molecular biology research. It contains the coding sequence for the PTEN (Phosphatase and Tensin Homolog) protein, a tumor suppressor gene. The vector can be used to express the PTEN protein in various cell lines for experimental purposes.
Automatically generated - may contain errors
2 protocols using pcdna pten
1
LPS-Induced H9C2 Cardiomyocyte Modeling
2
Investigating miR-425-5p and PTEN in Breast Cancer Cells
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Five BC cell lines (MCF-7, SKBR-3, MDA-MB-231, MDA-MB-468 and BT-20) and a normal human mammary epithelial cell line (MCF-10A) were obtained from the Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences). All cell lines were cultured in DMEM containing 10% fetal bovine serum (FBS) and incubated at 37°C in 5% CO2.
MCF-7 and MDA-MB-231 cells (3×105) were transfected with pre-miR-425-5p (miR-425-5p), mimic negative control (NC), miR-425-5p inhibitor, inhibitor NC or pcDNA-PTEN (Shanghai GenePharma Co., Ltd.). pcDNA plasmid without PTEN was used as a negative control. The sequences of the miRNA mimic, inhibitor and controls are presented inTable SI . The concentrations used for mimic, inhibitor and pcDNA were 50, 100 nM and 1000 ng/ul, respectively) Cells were transfected using Lipofectamine 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. Subsequent experiments were performed 24–72 h after transfection.
MCF-7 and MDA-MB-231 cells (3×105) were transfected with pre-miR-425-5p (miR-425-5p), mimic negative control (NC), miR-425-5p inhibitor, inhibitor NC or pcDNA-PTEN (Shanghai GenePharma Co., Ltd.). pcDNA plasmid without PTEN was used as a negative control. The sequences of the miRNA mimic, inhibitor and controls are presented in
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