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. 2020 Jun 19;8:513. doi: 10.3389/fcell.2020.00513

FIGURE 5.

FIGURE 5

miR-98 regulates HSCs activation depending on HLF/HIF-1α signaling pathway. (A) Schematic diagram of the putative HLF binding site within the human HIF-1α promoter. The LX-2 cells infected with Ad-HLF were subjected to ChIP assay with anti-Flag or IgG antibody. Representative results from three independent experiments are shown. (B) HIF-1α mRNA expression in activated LX-2 cells transfected with Ad-HLF or Ad-Con. (C) The luciferase reporter activity of HIF-1α promoter (HIF-1α-WT) or the mutant of HIF-1α promoter (HIF-1α-Mut) was measured in activated LX-2 cells infected with Ad-HLF or Ad-Con. (D) The correlation between HLF levels and HIF-1α expression in patient fibrotic liver tissues was assessed using Pearson’s correlation analysis, n = 33. (E) Proliferation of LX-2-pre-Sh-HLF or LX-2-pre-miR-98 cells transfected with Ad-HIF-1α or Ad-Con was detected by CCK8 assay. (F) The migration of the LX-2-pre-Sh-HLF or LX-2-pre-miR-98 cells transfected with Ad-HIF-1α or Ad-Con was compared using the Transwell assay, representative of three experiments. The number of cells was counted from different fields. (G) The expression levels of HLF, HIF-1α, TGF-β, p-Smad2, and p-Smad3 were examined in LX-2-pre-Sh-HLF or LX-2-pre-miR-98 cells transfected with Ad-HIF-1α or Ad-Con. Representative of three experiments. (H) The protein levels of α-SMA, Collagen-I, and TIMP-1 were examined by western blotting in LX-2-pre-Sh-HLF or LX-2-pre-miR-98 cells transfected with Ad-HIF-1α or Ad-Con. Representative of three experiments. Graph represents mean ± SEM.