Fig. 6

Hsa-miR-27b inhibits NF-кB signalling activity by targeting TAB3 in HCC cells. A GSEA of the correlation between hsa-miR-27b and NF-кB target gene signatures using the cohort of patients with HCC in the TCGA database. B NF-кB target gene mRNA (left) and protein (right) levels in the indicated cells. Data were normalized to GAPDH and are shown as fold change relative to the control cells for qRT-PCR (n = 3, Mann–Whitney test). GAPDH was used as a loading control for western blotting. C Dual luciferase reporter analysis for NF-кB transcriptional activity in the indicated cells. Data were normalized to the NC cells (n = 3, Mann–Whitney test). D Confocal images of immunofluorescence staining of p65 and p-p65 in the indicated cells. Scale bar, 200 μm (n = 3, Mann–Whitney test). E Representative IHC images and quantification of IHC scoring for p65 and phospho-p65 in subcutaneous tumours (n = 7, unpaired t-test). Scale bar, 200 μm. F Western blot analysis of protein expression in the indicated cells treated with TNF-α (10 ng/ml). GAPDH was used as a loading control. G Western blot analysis of p65 expression in the nucleic/cytoplasmic fractions after treatment with TNF-α (10 ng/ml) in the indicated cells. GAPDH or lamin A/C was used as a positive control for the cytoplasmic or nucleic fraction. B–G were repeated three times. *p < 0.05. **p < 0.01. Data are shown as mean ± SD. ES, enrichment score; NES, normalized enrichment score; NC, negative control; KD, knockdown; ctrl, control. CE, cytoplasmic extracts; NE, nuclear extracts