miRNA studies in in vitro and in vivo activated hepatic stellate cells

Figure 5 Changes during overexpression of rno-miR-146a in the hepatic stellate cell-2 cell line. A: Depicted are two putative binding sites of miR-146a to the 3’-UTR of rat tumor necrosis factor receptor associated factor 6 (TRAF6) and rat interleukin receptor associated kinase 1 (IRAK1), respectively; B: The Western blotting data show the suppression of TRAF6 and IRAK1, resulting in the decreased phosphorylation of IκB, although the expression of IκB remained unchanged. A representative Western blotting for two independent experiments is shown; C: Electrophoretic mobility shift assay (EMSA) results demonstrated a decrease in nuclear factor (NF)-κB DNA binding activity due to the overexpression of miR-146a. TATA binding protein (TBP) showed equal loading of samples. A representative EMSA experiment is shown out of three independent samples for each clone; D: miR-146a-overexpressing clones showed a reduced level of cyclooxygenase-2 (Cox-2) protein. The Western blotting shown is representative of two independent experiments; E: The relative fold change in mRNA expression between hepatic stellate cell (HSC)-2 and miR-146a-overexpressing HSC-2 cells for five different targets [NF-κB (RelA), Cox-2, smooth muscle α-actin, ColI, interleukin-6] is shown. The data represent the mean ± SE of two independent experiments (^aP≤ 0.005).