Figure 6.

Silencing HOTAIR Enhanced miR-29b Expression

(A) The levels of miR-29a, miR-29b, and miR-29c were analyzed in HSCs transfected with siHOTAIR. CCl₄ mice were treated with Ad-shHOTAIR. The levels of miR-29a, miR-29b, and miR-29c were detected in the livers from mice (B), as well as isolated primary HSCs (C), and primary hepatocytes (D). (E) Putative miR-29b binding sites (TS) within the mouse DNMT3b 3′-UTR are shown. The position of the binding sites was numbered relative to the first nucleotide of the 3′-UTR. Mutations were introduced into DNMT3b 3′-UTR that matched the seed region of miR-29b as shown in DNMT3b Mu. (F) Dual-luciferase assay was performed in HEK293T co-transfected with luciferase constructs containing the DNMT3b wild-type or Mu 3′-UTR and miR-29b mimics or scrambled oligonucleotides as the negative control. The mRNA (G) and protein (H) levels of DNMT3b in primary HSCs were reduced by miR-29b mimics, which were further decreased by siHOTAIR. *p < 0.05 compared to the control and ^#p < 0.05 compared to CCl₄ or miR-29b mimics group. Each value is the mean ± SD of three experiments.