PRDX2 shRNAs and scrambled control shRNA were purchased from GenePharma Company (Shanghai, China). Using a Lipofectamine 2000 (Invitrogen)-based transfection method, A549 cells were transfected with PRDX2 shRNAs or scrambled control shRNA and selected with puromycin for 3-4 weeks. Specifically, 293T cells (30% confluence) were cultured in 6-well plates and transfected with PRDX2 shRNAs and scrambled control shRNA for 48 hours. Then, the medium from 293T cells was collected and was added to A549 cells (30%-50% confluence) for 48 h, followed by selection with 6 μg/ml puromycin for 3-4 weeks. Plasmids (pcDNA3.1-PRDX2 and pcDNA3.1) were obtained from OBiO Technology Co. Ltd. (Shanghai, China). NCI-H1299 cells (70% confluence) were cultured in 6-well plates and transfected with pcDNA3.1-PRDX2 or pcDNA3.1 for 72 hours using Lipofectamine 2000.
Pcdna3
Manufactured by Obio Technology
Sourced in China
PcDNA3.1 is a mammalian expression vector used for the expression of recombinant proteins in cell lines. It contains the human cytomegalovirus (CMV) immediate-early promoter for high-level expression of the gene of interest, as well as an ampicillin resistance gene for selection in bacterial hosts.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using pcdna3
1
PRDX2 Knockdown and Overexpression in Lung Cancer Cells
2
Investigating miRNA-365 Regulation
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3
SPAG6 Knockdown and Overexpression in Cell Lines
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SPAG6 shRNA and scrambled control-shRNA were purchased from Shanghai GenePharma Company. For shRNA transfection, 293T cells were grown to 30–50% confluence in 6-well culture plates and transfected with scrambled shRNA or SPAG6 shRNA using Lipofectamine™ LTX reagent with PLUS™ reagent (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h. Then, the supernatant from 293T cells was harvested and added to Daudi and Raji cells for 72 h, followed by selection with 1 µg/ml puromycin for 1 month. Plasmids (pcDNA3.1-SPAG6 and pcDNA3.1) were purchased from Obio Technology Co., Ltd. For plasmid transfection, CA46 and NAMALWA cells (70% confluence) were plated in 6-well culture plates. Plasmids were purified and transfected with pcDNA3.1-SPAG6 or pcDNA3.1 for 72 h using Lipofectamine™ LTX reagent with PLUS™ reagent (Invitrogen; Thermo Fisher Scientific, Inc.), followed by selection with 1 µg/ml puromycin for 1 month.
4
Plasmid Transfection with TAK1 siRNA
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TAB1 plasmid and the vector pcDNA3.1(+) were purchased from Obio Technology Corp., Ltd. (Shanghai, China). SiRNAs of TAK1 and negative control (NC) were obtained from Beijing ViewSolid Biotech, the sequences are as follows: 5′-GGUGCUGAACCAUUGCCAUTT-3′ for si#1 of TAK1; 5′-GCAACCCAAAGCGCUAAUUTT-3′ for si#2 of TAK1; 5′-UUCUCCGAACGUGUCACGUTT-3′ for NC. Cells were transfected as previously described [25 (link)].
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