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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: HBx-induced MiR-1269b in NF-κB dependent manner upregulates cell division cycle 40 homolog (CDC40) to promote proliferation and migration in hepatoma cells

Fig. 2

NF-κB binds directly to the miR-1269b promoter and up-regulates its transcription. a The human miR-1269b genomic locus. The predicted promoter of miR-1269b, which contains two putative binding sites for NF-κB (pMiR-1269b-luc), and the mutant of miR-1269b promoter that does not contain NF-κB binding sites (pMiR-1269b-luc-M) are shown. b miR-1269b promoter-induced luciferase activity was increased in HepG2.2.15 cells compared to HepG2 cells. c The relative luciferase activity induced by the miR-1269b promoters constructed with or without NF-κB binding sites and the control vector in SMMC-7721 cells. d The effect of NF-κB (left) and TNF-α (right) on pMiR-1269b-luc and pMiR-1269b-luc-M in SMMC-7721 cells. e SMMC-7721 cells were co-transfected with pMiR-1269b-luc and pMiR-1269b-luc-M with puc18/HBV1.3, pFLAG/HBx(HBx) and the control vector, and luciferase activity was then detected after 24 h of transfection. f SMMC-7721 cells were cotransfected with pMiR-1269b-luc and pcDNA3 or HBx or NF-κB or both HBx and NF-κB. After 24 h, luciferase activity was analyzed. g EMSA to determine the DNA-binding activity of NF-κB using two substrates (probe 1 and probe 2). The results are shown as the mean ± SD of three experiments that were performed in duplicate. *p < 0.05, **p < 0.01

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