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. 2017 Jan 5;12(1):e0169702. doi: 10.1371/journal.pone.0169702

Fig 5. Effects of miR-223-3p overexpression on the human cells.

Fig 5

(A) Schematic diagram of the luciferase reporter plasmids including IL-17RD3’UTR wild type (WT) or deleted (del) miR-223-3p binding sequences in the 3’UTR in which six nucleotide forming the seed region of miR-223-3p were deleted. (B) Results of luciferase assay. Either pmG/hIl17rd WT or pmG/hIl17rd del was transfected into HeLa cells with either miR-223-3p overexpression vector (pBA/miR-223-3p) or negative control vector (pBA/NC). Results were expressed as the relative luciferase activity following transfection of cells with the negative control vector. *p<0.05 as compared with the pBA/NC group, #p<0.05 as compared with the pmG/hIL17rd WT group. (C) Results of miR-223-3p expression in MH7A cells. Results were expressed as the relative fold change following transfection of cells with the negative control vector. ***p < 0.001. (D) Results of Il17rd mRNA expression in MH7A cells. Results were expressed as the relative fold change following transfection of the negative control vector; ***p < 0.001. (E) Results of western blot analysis. (F) Relative expression of IL-17RD protein. IL17RD expression was normalized to that of β-actin in the same sample, and results were expressed as a percentage of the expression following transfection with the negative control vector. *p < 0.05. (G) Results of Il6 mRNA expression in MH7A cells. Results were expressed as the relative fold change following transfection with the negative control vector. ***p < 0.001. (H) Effect of miR-223-3p inhibitor on Il17rd mRNA expression. Results were expressed as the relative fold change with the negative control (NC). *p < 0.05. All results are presented as the means ± S.E. for each group (n = 3).