hsa-miR-409-3p regulates fibrinogen production via a target site in the FGB 3′-UTR. (A) Sequence details between the nucleotide 22 and 44 after the stop codon of the predicted target site, detected with TargetScan, for hsa-miR-409-3p in FGB 3′-UTR. (B) Normalized secreted fibrinogen levels for 2 control conditions: a negative control miRNA precursor (Neg. Ctrl.) and hsa-miR-144 (▭) and for hsa-miR-409-3p (). Precursors were transfected at 3 concentrations (3.3, 10, and 30nM), and secreted fibrinogen was analyzed. Nontransfected control (▬) was used to normalize results; n = 3. P values were calculated for experimental data versus values for Neg. Ctrl. (C) HEK-293T cells were cotransfected with firefly luciferase reporter plasmids carrying the FGB 3′-UTR (), the FGB-3′-UTR without the predicted target sequence of hsa-miR-409-3p (FGB-3′-UTR-Δseed, ▬), or a plasmid with no 3′-UTR (control plasmid, ▭); n = 5. P values were calculated for experimental data versus values for control plasmid conditions. (D) The relative quantity of each of the 5 fibrinogen transcripts quantified by quantitative RT-PCR from RNA samples of HuH7 cells transfected with 30nM of hsa-miR-409-3p, or hsa-miR-144 used here as a negative control; n = 3. P values were calculated for experimental data versus values for hsa-miR-144: *P < .05, **P < .01 (2-sided t test). Error bars represent SD. ns indicates not significant.