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. 2018 Oct 3;14(12):1732–1744. doi: 10.7150/ijbs.24232

Figure 5.

Figure 5

miR-7-5p specifically targeted p65. (A) The 3′-UTR of p65 contains a putative miR-7-5p binding site. The seed binding sequence of miR-7-5p in the WT 3′-UTR of p65 is indicated in red, while it is indicated in blue for mutant 3′-UTR. (B) The mutant of p65 3′-UTR failed to bind miR-7-5p. The miR-7-5p-mimic or its negative control (miR-NC) was co-transfected with pMIR-p65-3′-UTR-WT or pMIR-p65-3′-UTR-mutant into HFLS-RA cells for 24 h. The luciferase activity was quantified using the dual-luciferase assay reporter system by normalizing the signal to the Renilla activity. (C-F) The expression of miR-7-5p was negatively correlated with the p65 level. The miR-7-5p-mimic, anti-miR-5p or miR-NC was transfected into SW982 or HFLS-RA (HFLS) cells. The expression of miR-7-5p and p65 was determined in these cells. (G) The overexpression or downregulation of miR-7-5p affected the expression of NF-κB downstream targets. The cells used in C-F were subjected to qRT-PCR to examine the expression of NF-κB downstream targets, including TNF-α, IL-6, MCP-1 and CCL5. ** P < 0.001.