Human THP-1 cell line from Procell Life Science&Technology Co., Ltd. (Wuhan, China) was cultured in the RPMI-1640 medium supplied with 10% fetal bovine serum (FBS, Gibco, Gaithersburg, MD, USA) and 1% penicillin-streptomycin (Gibco) at 37°C with 5% CO2. Then, according to a previous study,36 (link) THP-1-derived macrophages were induced by treating with 50 ng/mL phorbol 12-myristate 13-acetate (PMA) for 48 h. Cell transfection was performed using Lipofectamine 3000 reagent (Invitrogen, USA) with the chemically synthesized circRNA small interfering RNAs (si-circ, sequence: 5ʹ-AGTACAACAATTCAGGTGATT-3ʹ), and small interfering RNAs negative control (si-NC, sequence: 5ʹ-UUCUCCGAACGUGUCACGUTT-3ʹ). After 48 h of transfection, the macrophages were treated with type I interferon (IFN-I, 1000 units/mL, PBL Assay Science, USA) to induce the inflammation.
Protocol detail
THP-1 Macrophage Inflammation Induction
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Assay
Cell
Cell culture
Circrna
Cultured medium
Human
Ifn i
Inflammation
Lipofectamine
Macrophages
Penicillin
Phorbol myristate acetate
Small interfering rnas
Streptomycin
Thp 1 cell line
Transfection
Corresponding Organization : University of Freiburg
Other organizations : Xiangya Hospital Central South University, Central South University
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Variable analysis
independent variables
- Treatment with 50 ng/mL phorbol 12-myristate 13-acetate (PMA) for 48 h to induce THP-1-derived macrophages
- Transfection with chemically synthesized circRNA small interfering RNAs (si-circ)
- Transfection with small interfering RNAs negative control (si-NC)
- Treatment with type I interferon (IFN-I, 1000 units/mL) to induce inflammation
- Unspecified
- Culturing THP-1 cells in RPMI-1640 medium with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C with 5% CO2
- Positive control: Treatment with type I interferon (IFN-I) to induce inflammation
- Negative control: Transfection with small interfering RNAs negative control (si-NC)
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