Journal: European Journal of Medical Research

Article Title: MiRNA-29b accelerates the PDGF in exosomes and stimulates hepatic stellate cells to promote liver fibrosis in biliary atresia

doi: 10.1186/s40001-025-02731-z

Figure Lengend Snippet: Effects of miRNA-29b overexpression and inhibition in exosomes on the expression of DNMT3a, DNMT3b, PDGFA, COL1A1, and α-SMA in LX-2 cells and verification of miRNA-29b target genes by dual luciferase assays. a miRNA-29b-overexpressing exosomes were cocultured with LX-2 cells, and the expression levels of DNMT3a and DNMT3b in the Up group were significantly decreased. b The transcription level of PDGFA in the b Up group was significantly increased. c The expression levels of COL1A1 and α-SMA in the Up group were increased. d Exosomes with low miRNA-29b expression were cocultured with LX-2 cells, and the expression levels of DNMT3a and DNMT3b in the Down group were significantly increased. e The expression level of PDGFA in the Down group was decreased. f The expression of COL1A1 and α-SMA decreased in the Down group. g Double luciferase experiments proved that DNMT3a and DNMT3b were the target genes of miR-29b-3p. Note: Normal: untreated LX-2 cells in the blank control group; control: negative empty exosomes were cocultured with LX-2 cells. Down: exosomes with low miRNA-29b expression were cocultured with LX-2 cells; a–f : LX-2 cells cocultured with exosomes from 3 independent experiments (n = 3); g: Data from 4 biological replicates (n = 4). * indicates p < 0.05 compared with the control group

Article Snippet: The membrane was blocked with 5% skim milk and incubated with primary antibodies against DNMT3a, DNMT3b, PDGFA, COL1A1 and α-SMA (GeneTex, Abcam).

Techniques: Over Expression, Inhibition, Expressing, Luciferase, Control

Journal: European Journal of Medical Research

Article Title: MiRNA-29b accelerates the PDGF in exosomes and stimulates hepatic stellate cells to promote liver fibrosis in biliary atresia

doi: 10.1186/s40001-025-02731-z

Figure Lengend Snippet: Effect of 5-AZAC on LX-2 cells cocultured with miRNA-29b overexpression and inhibition exosomes. a The miRNA-29b overexpression exosome + 5Azac group showed decreased expression of DNMT3a and DNMT3b; b the miRNA-29b overexpression exosome + 5Azac group showed increased expression of PDGFA; c The expression of COL1A1 and α-SMA in the miRNA-29b overexpressed exosome + 5Azac group increased; d The expression levels of DNMT3a and DNMT3b in the miRNA-29b low-expression exosome + 5Azac group were decreased; e the miRNA-29b low expression exosome + 5Azac group showed increased expression of PDGFA; f The expression of COL1A1 and α-SMA increased in the miRNA-29b low-expression exosome + 5Azac group. Note : Normal: untreated LX-2 cells in the blank control group; control: negative empty exosomes were cocultured with LX-2 cells. Up + 5Azac: miRNA-29b-overexpressing exosomes + 5Azac were cocultured with LX-2 cells. Down + 5Azac: Exosomes with low expression of miRNA-29b + 5Azac were cocultured with LX-2 cells; a – f : Data from 3 independent experiments (n = 3). * indicates p < 0.05 compared with the control group

Article Snippet: The membrane was blocked with 5% skim milk and incubated with primary antibodies against DNMT3a, DNMT3b, PDGFA, COL1A1 and α-SMA (GeneTex, Abcam).

Techniques: Over Expression, Inhibition, Expressing, Control

Journal: European Journal of Medical Research

Article Title: MiRNA-29b accelerates the PDGF in exosomes and stimulates hepatic stellate cells to promote liver fibrosis in biliary atresia

doi: 10.1186/s40001-025-02731-z

Figure Lengend Snippet: Detection of the relative expression of transcript level and protein level of each gene in UP group and Up + 5Azac group, Down group and Down + 5Azac group. a–b: At both transcriptional and protein levels, the Up + 5Azac group showed significantly lower DNMT3a/3b enzyme activities, higher PDGFA methylation levels, and increased COL1A1 and α-SMA expression compared to the Up group. c-d: At the transcriptional and protein levels, the Down + 5Azac group showed decreased DNMT3a/3b enzyme activity, increased PDGFA methylation levels, and increased COL1A1 and α-SMA expression compared to the Down group. Note: Up: exosomes overexpressing miRNA-29b were cocultured with LX-2 cells; Up + 5Azac: miRNA-29b -overexpressing exosomes + 5Azac were cocultured with LX-2 cells; Down: exosomes with low miRNA-29b expression were cocultured with LX-2 cells; Down + 5Azac: Exosomes with low expression of miRNA-29b + 5Azac were cocultured with LX-2 cells. a – d : LX-2 cells cocultured with exosomes from 6 independent experiments (n = 6). * indicates p < 0.05; ** indicates p < 0.01; *** indicates p < 0.001; **** indicates p < 0.0001

Article Snippet: The membrane was blocked with 5% skim milk and incubated with primary antibodies against DNMT3a, DNMT3b, PDGFA, COL1A1 and α-SMA (GeneTex, Abcam).

Techniques: Expressing, Methylation, Activity Assay