Skip to main content
. 2012 Sep 6;2:637. doi: 10.1038/srep00637

Figure 2. MiR122 modulates ISRE activities.

Figure 2

(a) Overexpression of the selected microRNA precursors suppresses ISRE activity following IFN-α stimulation. ISRE reporter plasmids were transiently transfected, with or without selected microRNA precursor-expressing plasmids, into Huh7 cells. Luciferase values were normalized to those of cells transfected with an empty vector and without IFN, which were set to 1. *, p < 0.05. Data represent the means ± standard deviations (SD) of three independent determinations. Similar results were obtained using HepG2 cells. (b) p53 activities were unaffected by miR122 expression. Reporter assays were performed with p53 reporter and p53 expressing plasmids with or without miR122 precursor expression in Huh7 cells. Data represent the means ± SD of three independent determinations. (c) Silencing of miR122 function enhances ISRE activity. Anti-miR122 expressing plasmids were used to perform an ISRE reporter assay in Huh7 cells. *, p < 0.05. Data represent the means ± SD of three independent determinations. (d) Hela-Tet-Off-miR122 precursor cell lines were used to measure ISRE activity. Cells were transfected with ISRE reporter constructs with (DOX+) or without (DOX-) doxycyclin. DOX+ shut off the miR122 expression in these cells. The reporter activities after 6 h of IFN-α stimulation were compared. *, p < 0.05. Data represent the means ± SD of three independent determinations. (e, f) IFN-inducible genes (e, ISG15; f, IFNAR1) were more induced in miR122-silenced Huh7 cells, determined by quantitative RT-PCR using RNA after 6 h of IFN-α stimulation. *, p < 0.05. Data represent the means ± SD of three independent determinations.