Research Article

Mmu-miR-27a-5p-Dependent Upregulation of MCPIP1 Inhibits the Inflammatory Response in LPS-Induced RAW264.7 Macrophage Cells

Figure 3

Mmu-miR-27a-5p targets MCPIP1. (a) RAW264.7 were transfected with a luciferase construct containing MCPIP1 3′-UTR (MCPIP1 -UTR-luc) and GAPDH 3′-UTR (GAPDH-UTR-luc) in the presence of mmu-miR-27a-5p. (b) Putative binding site of mmu-miR-27a-5p in MCPIP1 3′-UTR. (c) Firefly luciferase activity of the deletion mutant of predicted target sites. Firefly luciferase activity of HEK293 cells cotransfected with Pre-miR-NC (negative control, white bars) or Pre-miR-27a-5p (gray bars) plus pmir-luciferase-MCPIP1-3′-UTR (lane 1) and pmir-luciferase-MCPIP1-3′UTR-mut (lane 2), respectively. Results depict the average (±SEM) of three independent transfections. (d) A series of reporter plasmids was constructed for luciferase assays. (e) Firefly luciferase activity of HEK293 cells cotransfected with Pre-miR-NC (negative control, white bars) or Pre-miR-27a-5p (gray bars) plus pmir-luciferase-MCPIP1-3′UTR-240 (lane 1), pmir-luciferase-MCPIP1-3′UTR-410 (lane 2), pmir-luciferase-MCPIP1-3′UTR-567 (lane 3), and pmir-luciferase-MCPIP1-3′UTR-810 (lane 4).
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