Fig. 2: miR-139-5p is a target of NEAT1. | Cell Death Discovery

Fig. 2: miR-139-5p is a target of NEAT1.

From: miR-139-5p sponged by LncRNA NEAT1 regulates liver fibrosis via targeting β-catenin/SOX9/TGF-β1 pathway

Fig. 2

A A schematic diagram demonstrated the putative sites of miR-139-5p binding with NEAT1. B Relative luciferase activities of pmirGLO-NEAT1-WT or pmirGLO-NEAT1-MUT were detected in HEK293T cells. C RIP experiments were carried out with Ago2 antibody on extracts from LX-2 cells. Relative expression level of NEAT1 was shown as fold enrichment in Ago2 relative to IgG immunoprecipitates by qRT-PCR. D Interaction between NEAT1 and miR-139-5p was verified by pull-down assay. n = 3/group. E RNA in situ hybridization was performed to observe the co-localization between miR-139-5p and NEAT1. n = 3/group. Next, LX-2 cells were transfected with Ad-NEAT1 or Ad-shNEAT1 for 48 h. F qRT-PCR was performed to detect the expression level of miR-139-5p in LX-2 cells transfected with Ad-NEAT1 or Ad-shNEAT1. n = 3/group. G The protein levels of α-SMA, Collagen-I, and TIMP-1 in LX-2 cells transfected with Ad-NEAT1 or Ad-shNEAT1 were detected by western blotting. Representative of three experiments. H Confocal laser microscopy was used to analyze the immunofluorescence staining for α-SMA (green) in LX-2 cells after transfection with Ad-NEAT1 or Ad-shNEAT1. DAPI was used to stain nuclei, blue. Original magnification×400. Representative of three experiments. I Flow cytometry was performed to analyze the cell-cycle distribution of LX-2 cells transfected with Ad-NEAT1 or Ad-shNEAT1. Representative of three experiments. J CCK8 assay was performed to detect the proliferation of LX-2 cells transfected with Ad-NEAT1 or Ad-shNEAT1. K Flow cytometry was performed to detect the cell apoptosis of LX-2 cells transfected with Ad-NEAT1 or Ad-shNEAT1. Representative of three experiments. L The protein levels of α-SMA, Collagen-I, and TIMP-1 in LX-2 cells transfected with Ad-NEAT1 and miR-139-5p were detected by western blotting. Representative of three experiments. M Confocal laser microscopy was used to analyze the immunofluorescence staining for α-SMA (green) in LX-2 cells after transfection with Ad-NEAT1 and miR-139-5p. DAPI was used to stain nuclei, blue. Original magnification×400. Representative of three experiments. N CCK8 assay was performed to detect the proliferation of LX-2 cells transfected with Ad-NEAT1 and miR-139-5p. Graph represents mean ± SD. *P < 0.05, **P < 0.01.

Back to article page