At 24 h following transfection, THP-1 cells were washed with Hank's balanced salt solution (Beijing Solarbio Science & Technology Co., Ltd.) before the culture medium was replaced with fresh RPMI-1640 medium. A final concentration of 1 µg/ml LPS (Sigma-Aldrich; Merck KGaA) was subsequently added into the culture medium, followed by incubation for a further 3 h. The cells were then collected for the measurement of IRAK1 and p68 expression, whilst the supernatant of the culture medium was collected for the measurement of cytokine levels, as described previously (15 (link)). The cells were divided into the following experimental groups: Blank control, no RNA transfection or LPS treatment; LPS group, treated with LPS only; mimic group, miR-146 amimic+LPS; NC mimic group, NC mimic + LPS; inhibitor group, miR-146a inhibitor + LPS; and inhibitor NC group, inhibitor NC + LPS.
Protocol detail
Modulation of IRAK1 and p68 Expression in LPS-Stimulated THP-1 Cells
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Corresponding Organization :
Other organizations : Tianjin First Center Hospital, Tianjin Medical University, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin Children's Hospital
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Variable analysis
independent variables
- Transfection with different RNA molecules (blank control, miR-146a mimic, NC mimic, miR-146a inhibitor, inhibitor NC)
- Lipopolysaccharide (LPS) treatment
- IRAK1 expression
- P68 expression
- Cytokine levels in the culture medium
- Cell line (THP-1 cells)
- Culture medium (RPMI-1640)
- Washing solution (Hank's balanced salt solution)
- Incubation time (3 h after LPS treatment)
- Blank control (no RNA transfection or LPS treatment)
- LPS group (treated with LPS only)
- NC mimic group (NC mimic + LPS)
- Inhibitor NC group (inhibitor NC + LPS)
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