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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: miR-203, fine-tunning neuroinflammation by juggling different components of NF‐κB signaling

Fig. 1

miRNA-203 targeted downregulation of Akirin2 expression in vivo and vitro. A The conserved miRNA:mRNA interaction site in the 3ʹUTR of Akirin2 predicted using TargetScan. B Luciferase assay showed a direct sequence-specific targeting of miR-203 on Akirin2 3ʹUTR. The results were presented as means of normalized luciferase activities of three biological replicated each measured in triplicates, with error bars showing the standard deviation. Asterisks indicate statistical significance (ANOVA, Turkey’s post hoc). C and D The 217 bp Akirin2 3ʹUTR containing human wild-type (C) or sense mutation in the seed sequence of the miR-203 binding site (D) were cloned into pmiRGLO and co-transfected into 293T cells with miRNA-203 mimics or nonsense scramble RNA, respectively. Luciferase activities were measured 48 h post transfection. E and H Downregulated Akirin2 expression was detected 48 h post-transfection using western blots in cultured BV2 cells transfected with miRNA-203 mimics or scramble RNA, respectively. β-Actin was used as an internal control. Luminescence-based relative quantification of protein (Akirin2:β-actin) of three individual biological replicates with error bars representing the standard deviation showed significant decreasing of Akirin2 expression in cells transfected with miR-203 mimics (ANOVA, Turkey’s post hoc). F and I Intrinsic expression of Akrin2 was decreased in mouse hippocampus with stereotactic injection of GV309-pre-miR203-eGFP, comparing with that in control mice injected with GV309-Scr-eGFP. Results were presented as the luminescence-based relative quantification of western blots of protein (Akirin2:β-actin) of three individual biological replicates with error bars showing the standard deviation. G and J Intrinsic expression of 14-3-3θ was decreased in mouse hippocampus with stereotactic injection of GV309-pre-miR203-eGFP, comparing with that in control mice injected with GV309-Scr-eGFP. Results were presented as the luminescence-based relative quantification of western blots of protein (14-3-3θ:β-actin) of three individual biological replicates with error bars showing the standard deviation. Asterisks indicate statistical significance between samples (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)

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