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)
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Sirt1 Overexpression in Insulin-Resistant Cells
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Corresponding Organization : Yixing People's Hospital
Other organizations : First People’s Hospital of Jingmen
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Variable analysis
independent variables
- Overexpression of Sirt1 using the pcDNA-Sirt1 plasmid
- Addition of the IRS1 inhibitor NT157
- Not explicitly mentioned
- Cell density (2×10^5 cells/mL)
- Incubation of the plasmids (pcDNA-Sirt1 and pcDNA-NC) in Opti-MEM using Lipofectamine 2000 according to the manufacturer's protocol
- Negative control (pcDNA-NC)
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