Human embryonic liver cell line L02 was purchased from China Center for Type Culture Collection (Wuhan, China). N-acetyl-L-cysteine (NAC) (Beyotime, Shanghai, China) (5 mmol/L)[22 (link)], nicotinamide (NAM) (Beyotime) (5 mmol/L)[23 (link)], GW6471 (Sigma-Aldrich) (3 μM)[24 (link)] or Compound C (Sigma-Aldrich) (10 μM)[25 (link)], which were dissolved in dimethyl sulfoxide (Sigma-Aldrich), were used to pretreat L02 cells for 1 h, followed by LPS (5 μg/mL)[26 (link)] treatment. Hypoxic conditions (1% O2) were obtained using humidified variable aerobic workstation InVivo2 400 (Ruskinn, Pencoed, United Kingdom)[27 (link)]. For transient transfection, cells were transfected with 2 μg plasmid of pECE-flag-Sirt1 (Addgene, Cambridge, MA, United States) and pECE empty vector (Addgene).
Protocol detail
Human Liver Cell Line L02 Response
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N-acetyl-L-cysteine (NAC)
nicotinamide
GW6471
Compound C
dimethyl sulfoxide
pECE-flag-Sirt1
pECE empty vector
Aerobic
Cell line
Cells
Dimethyl sulfoxide
Embryonic
Gw6471
Human
Hypoxic
Liver
N acetyl l cysteine
Nicotinamide
Plasmid
Sirt1
Transfection
Transient
Vector
Corresponding Organization :
Other organizations : Renmin Hospital of Wuhan University, Wuhan University
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Variable analysis
independent variables
- N-acetyl-L-cysteine (NAC) (5 mmol/L)
- Nicotinamide (NAM) (5 mmol/L)
- GW6471 (3 μM)
- Compound C (10 μM)
- LPS (5 μg/mL)
- Hypoxic conditions (1% O2)
- Not explicitly mentioned
- Dimethyl sulfoxide (Sigma-Aldrich) used to dissolve the compounds
- PECE empty vector (Addgene) used for transient transfection
- Not mentioned
- PECE empty vector (Addgene) used for transient transfection
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