Additional file 7: of MicroRNA miR-29 controls a compensatory response to limit neuronal iron accumulation during adult life and aging

Mir-29 target D. rerio and M. musculus Ireb2 3′-UTR. A Expression of GFP assessed by cytofluorometric analysis. Fusion with Ireb2 3′-UTR of D. rerio. The gray column indicates the baseline fluorescence intensity of the construct without miR-29 mimic. The middle blue column indicates the fluorescence with miR-29 mimic and the red column the fluorescence of a construct (Δ) where the putative binding site for miR-29 in the Ireb2 3′-UTR was mutated to destroy complementarity. Statistical significance of fluorescence difference between baseline and co-injection with miR-29 mimics was evaluated by Student’s t-test (** P < 0.01). B 293 T cells co-transfected with renilla and firefly luciferase construct containing the wild-type or the mutant target sites of mmu-Ireb2 3′-UTR along with the miRNA expression plasmid (pcs2+: CMV:RFP-miR-29b/c precursor-polyA-tail) or the empty vector (pcs2+: CMV:RFP-polyA-tail). Histograms show normalized (to renilla) sensor luciferase activity of empty vector transfected cells (gray column) respect to transfected cells with miRNA expressing vector (blue column) or to mutated sensor with miRNA expressing vector. Bars represent mean ± standard deviation (Student’s t-test, **P < 0.01) derived from two independent experiment performed in triplicates for each condition. (PDF 41 kb