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

Journal: Hepatology Communications

Article Title: Single-cell transcription reveals hepatocyte-to-cholangiocyte reprogramming and biliary gene profile in biliary atresia

doi: 10.1097/HC9.0000000000000710

Figure Lengend Snippet: Enriched pathways and genes in the cholangiocytes of BA. (A) Flowchart of isolating BA epithelial cells from the merged groups. (B) t-SNE visualization of hep, rep, and cho in BA. (C) Violin plots showing the expression levels of the marker genes of hep, rep, and cho in BA. (D) Heatmap displaying enriched pathways of cholangiocytes in BA by GSVA analysis. (E) Violin plots displayed the EMT score among the 3 types of epithelial cells in BA. (F) Heatmap displaying the average expression level of EMT markers, fibrotic markers, inflammatory genes, and m 6 A readers in BA hep, rep, and cho. (G) Representative immunohistochemistry sections of CDH2, smad2/3, FGFR2, PDGFA, CXCL6, and IGF2BP2 in a TMA. Dashed circles in PDGFA were recognized as cholangiocytes. H. Representative immunofluorescence staining of CDH2, Smad2/3, FGFR2, PDGFA, CXCL6, and IGF2BP2 in TMA of BA group. I. Box plots showing the expression levels of the indicated genes in TMA, colored by different groups. * p <0.05; ** p <0.01; *** p <0.001; **** p <0.0001. Abbreviations: BA, biliary atresia; CHO, cholangiocytes; CS, cholestasis; EMT, epithelial-mesenchymal transition; HEP, hepatocytes; NC, normal control; REP, reprogrammed cells; TMA, tissue microarray.

Article Snippet: After antigen blocking with 5% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: CK19 (Abcam, ab52625, 1:600), HNF4A (Abcam, ab201460, 1:2000), SOX9 (CST, 82630, 1:600), CDH2 (CST, 13116, 1:200), Smad2/3 (CST, 8685, 1:400), FGFR2 (CST, 23328, 1:200), PDGFA (santa cruz, sc-9974, 1:200), CXCL6 (CST, 73201, 1:100), IGF2BP2 (Abcam, ab124930, 1:200), CFTR (Abcam, ab270238, 1:500), MMP7 (Abcam, ab207299, 1:4000), VTCN1 (Abcam, ab209242, 1:1000), LAMC2 (Abcam, ab210959, 1:500), ANKRD1 (Proteintech, 11427-1-AP, 1:200), ELF3 (Sigma, HPA003479, 1:200), KLF5 (Abcam, ab273672, 1:500), and HNF1B (Abcam, ab128912, 1:100).

Techniques: Expressing, Marker, Immunohistochemistry, Immunofluorescence, Staining, Control, Microarray