FIGURE. 2.

miR-31 regulation TNF-induced E-selectin expression by targeting the E-selectin 3’UTR. COS-7 cells were co-transected with the indicated constructs and with 40 nM of M-miR-31 or CM (A) or with SELE 3’UTR construct and 40 nM of the indicated miRNA mimics or CM (B). Data are expressed as relative luciferase activity to control samples co-transfected with an equal concentration CM (mean ± SEM of three experiments performed in duplicate). *Significantly different from cells transfected with CM and with control 3’UTR and with the indicated miRNAs, p≤0.05. C and D, Western blot analysis of E-selectin protein levels in ECs, (C) transfected for 12 h with M-miR-31 or CM or (D) with I-miR-31 or CI, in both cases 24 h post-transfection the cells were treated or not with TNF. Hsp-90 was used as loading control. VE-cadherin served as a control protein that is not targeted by miR-31. Graphs in the right show the relative total E-selectin protein levels compared to non-treated CM or CI-transfected controls (mean ± SEM of four experiments). *Significantly different from TNF treated control, p≤0.05.