Skip to main content
. 2017 Jul 27;13(7):e1006896. doi: 10.1371/journal.pgen.1006896

Fig 4. miR-1254 down-regulates HO-1 transcription via targeting TFAP2A.

Fig 4

(A) qRT-PCR analysis of TFAP2A, USF1 and NF-κB mRNA levels in cells transfected with indicated oligonucleotides. (B) Luciferase activity assays of the PGL-HO1 reporter in HEK293 cells with TFAP2A knockdown (left) or over-expression (right). (C) Western blot analysis of the effect of TFAP2A knockdown on HO-1 protein expression compared with miR-1254 in A549 cells. (D) The mRNA levels of TFAP2A, USF1 and NF-κB in CRISPR/Cas9-modified miR-1254 knockdown A549 cells. (E) Western blot analysis of the protein level of TFAP2A in the CRISPR/Cas9-modified miR-1254 knockdown A549 cells. (F) HO-1 protein levels in cells transfected with the indicated oligonucleotides (miR-1254 and its mutants) for 48 h assayed by immunoblotting. Left: Representative images. Right: Statistical results. (G) Western blot analysis of the TFAP2A over-expression effect on HO-1 protein expression compared with miR-1254 in A549 cells. (H) ChIP analysis of the polⅡ and TFAP2A enrichment in HO-1 promoter in A549 cells using antibodies against polⅡand TFAP2A, with IgG as a negative control. Left: Representative images. Right: Statistical results. Data are presented as the mean ± SEM of three independent experiments. *P<0.05, **P and ***P <0.01 vs. nc; #P<0.05, ##P and ###P <0.01 vs miR-1254.