LOVO and SW480 cells (1 × 105) were seeded onto 6-well plates. Short interfering RNA of SIRT1 (siSIRT1) 5′- CCAAGCAGCUAAGAGUAAUTT-3′ (sense), 5′- AUUACUCUUAGCUGCUUGGTT-3′ (antisense) (Genepharma, China) and pcDNA-SIRT1 (Genepharma, China) were transiently transfected into LOVO and SW480 cells when cells reached 70–80% confluence using Lipofectamine™ 3000 Reagent (Invitrogen, USA). After 48 h transfection, the cells were collected for further experiments.
Pcdna sirt1
Manufactured by GenePharma
Sourced in China
PcDNA-SIRT1 is a plasmid vector that contains the coding sequence for the SIRT1 gene. SIRT1 is a protein that plays a role in cellular processes such as gene expression, metabolism, and longevity. The PcDNA-SIRT1 vector can be used for the expression and study of the SIRT1 protein in various cell lines and research applications.
Automatically generated - may contain errors
3 protocols using pcdna sirt1
1
Modulating SIRT1 expression in colorectal cancer
2
Sirt1 Overexpression in Insulin-Resistant Cells
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The insulin-resistant FL62891 and HHL5 cells were seeded at a density of 2×105 cells/mL in 6-well plates. The plasmid of Sirt1 was used for Sirt1 overexpression. Then, the overexpression plasmid of Sirt1 (pcDNA-Sirt1, 500ng/µl, GenePharma, Shanghai, China) and negative control (pcDNA-NC, GenePharma) were incubated in Opti-MEM (Invitrogen, Carlsbad, USA) using LipofectamineTM 2000 (Invitrogen, Carlsbad, USA) according to the manufacturer’s protocol. The inhibitor NT157 of IRS1 (3 μM, Beijing Bio Lebo Technology Co., Ltd, Beijing, China) was added in FL62891 and HHL5 cells.15 (link)
3
Regulation of Breast Cancer by miR-22 and SIRT1
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miR-22 mimics (miR-22), miR-22 inhibitor (anti-miR-22), scrambled negative control miRNA (miR-NC), siRNA specific targeting sirt1 (si-sirt1), scrambled negative control siRNA (si-NC), and plasmid encoding sirt1 (pcDNA-sirt1) were all synthesized by GenePharma Co. Ltd. (Shanghai, China). Breast cancer cells (1 × 105) were plated into 6-well plates and cultured in medium without antibiotics for approximately 24 h prior to transfection. The next day, cells were transiently transfected with miRNAs, siRNAs or plasmids using Lipofectamine 2000 (Invitrogen). Cells were collected 48 h post-transfection for functional analysis.
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