Fig. 5: miR-139-5p regulates HSCs activation depending on the interaction between β-catenin and SOX9.

A The mRNA expression level of SOX9 was upregulated in human fibrotic liver tissues. n = 30/group. B The sections of human liver tissues were treated with SOX9 immunohistochemical staining, n = 6/group, original magnification×200. C Predicted protein network visualization with STRING. The network nodes represented proteins. These proteins were clustered with k-means clustering algorithms. D Co-immunoprecipitation assays revealed that β-catenin could bind with SOX9 in activated LX-2 cells. E Moreover, Dual immunofluorescence staining of activated LX-2 cells using anti-β-catenin and anti-SOX9 antibodies was performed. Representative of three experiments. DAPI was used to stain the nuclei, original magnification×400. F The protein levels of SOX9, α-SMA, Collagen-I, and TIMP-1 in LX-2-pre-miR-139-5p cells infected with Ad-SOX9 were detected by western blotting. Representative of three experiments. G Immunofluorescence staining for α-SMA (green) in LX-2-pre-miR-139-5p cells infected with Ad-SOX9 was analyzed by confocal laser microscopy. DAPI was used to stain the nuclei, blue. Original magnification×400, Representative of three experiments. H Proliferation of LX-2-pre-miR-139-5p cells infected with Ad-SOX9 was analyzed by CCK8 assay. I The wound-healing assay of LX-2-pre-miR-139-5p cells infected with Ad-SOX9 or Ad-Con was performed to observe the migration capability. Representative of three experiments. J The transwell assay of the LX-2-pre-miR-139-5p cells infected with Ad-Ad-SOX9 or Ad-Con was performed to analyze the migration capability, representative of three experiments. The number of cells was calculated from different fields. Graph represents mean ± SD. *P < 0.05, **P < 0.01.