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. 2017 Dec 18;9(12):1370. doi: 10.3390/nu9121370

Figure 5.

Figure 5

FOXO3 is a target of miR-132. (A) miR-132 targets were predicted using TargetScan (www.targetscan.org) and microRNA (www.microrna.org); (B) PA induces the mRNA expression of FOXO3; (C) PA induces the protein expression of FOXO3; (D–G) THP-1 cells were transfected with a mimic negative control, mimic, inhibitor negative control, or inhibitor of miR-132 at 20 nM for 48 h before treatment with 200 μM PA for 12 h; (D,F) The effect of the miR-132 mimic and inhibitor on the FOXO3 mRNA content was determined by quantitative RT-PCR; (E,G) Cell lysates were subjected to Western blot analysis using antibodies specific for FOXO3 and β-actin. β-Actin was used as a loading control; (H) The effect of FOXO3 silencing on miR-132 expression was determined by quantitative RT-PCR in THP-1 cells. NS, no significance; (I,J) Primary murine macrophages were treated with 200 μM PA for 12 h. Levels of FOXO3 mRNA and protein expression were measured; (K) Primary murine macrophages were transfected with inhibitor negative control or inhibitor of miR-132 at 20 nM for 48 h before being treated with 200 μM PA for 12 h. FOXO3 mRNA was examined by quantitative RT-PCR. # p < 0.05 compared with the mimic or inhibitor negative control in PA-untreated cells, and * p < 0.05 compared with the mimic or inhibitor negative control in PA-treated cells.