Fig. 6

BRF2 knockdown changes key proteins within the Wnt/β-catenin pathway. Bioinformatic analysis indicates that the Wnt/β-catenin pathway may be a downstream signaling pathway of BRF2. A Volcano plot of single gene difference analysis according to differential BRF2 expression according to TCGA liver hepatocellular carcinoma (LIHC) database. Fold change ≥ 1.5; P < 0.05. B Representative graphs of KEGG/GO pathway analysis for the differentially expressed genes in different BRF2 expression groups. C Left: Western blots were used to verify the expression of key proteins in the downstream pathway after knockdown of BRF2. Right: Quantitative analysis of key proteins. ***, P < 0.001. D Representative western blot data showing N-cadherin and β-catenin in Huh-7 cells and Smmc-7721 cells treated by OE-BRF2 and NC. E Representative western blot data showing N-cadherin and β-catenin in Huh-7 cells and Smmc-7721 cells after infection with OE-BRF2 and OE-BRF2 + LGK974. F, G Invasion and migration of Huh-7 cells and Smmc-7721 cells treated by OE-BRF2 and OE-BRF2 + LGK974 were measured by wound-healing assay. Left: Representative images at 100 × magnification; scale bar, 100 µm. Right: Quantification of wound-healing assay. ∗  ∗  ∗ , P < 0.001, OE-BRF2 vs OE-BRF2 + LGK974 group. H, I Invasion and migration of Huh-7 cells and Smmc-7721 cells treated by OE-BRF2 and OE-BRF2 + LGK974 were measured by transwell migration and invasion assays. Left: Representative images at 100 × magnification; scale bar, 100 µm. Right: Quantification of transwell migration and invasion assays. ∗  ∗  ∗ , P < 0.001, OE-BRF2 vs OE-BRF2 + LGK974 group. J, K Left: Representative images of wound-healing assay in Huh-7 cells and Smmc-7721 cells treated by OE-BRF2 and NC. Magnification, 100 × ; scale bar, 50 µm. Right, Quantification of wound-healing assay. n = 3. ∗ ∗  ∗ , P < 0.001. OE-BRF2 vs NC group