Journal: Nature Communications

Article Title: Cellular crosstalk mediated by TGF-β drives epithelial-mesenchymal transition in patient-derived multi-compartment biliary organoids

doi: 10.1038/s41467-025-61442-5

Figure Lengend Snippet: a Overview of in vitro A8301-mediated TGF-β signal inhibition in BA-MBOs. Created in BioRender. Ayabe, H. (2025) https://BioRender.com/q1pk62z . b Representative plots and quantification of FACS of day 14 MBOs treated with A8301. Number of hybrid (pan-CK + /VIM + ), cholangiocyte (pan-CK + /VIM - ) and mesenchymal (pan-CK − /VIM + ) cells were quantified (Mean ± SD; n = 12 organoids). c Images of MBOs treated with A8301 captured before collection and after whole-mount immunostaining (CK19: red, CD31: green, VIM: white). Abundance of CK19 + structures (Mean ± SD; n = 9 organoids examined over 3 independent experiments) and size distribution of PBG-like structures were quantified ( n = 39-104 clusters examined over 3 independent experiments). d Immunostaining of ECAD (green), CK19 (red) and Nucleus (blue; DAPI) in MBOs treated with A8301. ECAD + cells in CK19 + cells in the field were quantified (Mean ± SD; n = 12 fields from 3 organoids). e Immunostaining of ZO1 (green), CK19 (red) and Nucleus (blue; DAPI) in MBO treated with A8301. ZO1 + cells in CK19 + cells per field were quantified (Mean ± SD; n = 12 fields from 3 organoids). f Alcian blue staining of MBOs treated with A8301. Mucin production was determined by ratio of Alcian Blue + areas in CK19 + (from separately examined immunofluorescence data) areas (Mean ± SD; n = 12 fields from 3 organoids). g Top enriched terms of MSigDB Hallmark 2020 , and GO , sets in serum proteins (reanalyzed from Lertudomphonwanit et al. ) upregulated in BA subjects ( n = 175 patients) vs age-matched normal controls ( n = 9). MF: Molecular Function, BP: Biological Process. h Heatmap of enriched proteins in the EMT pathway in MSigDB Hallmark 2020 sets. ( n = 9 NC and 175 BA subjects). i Serum TGF-β/Activin ligand levels represented by the relative fluorescent unit (RFU; Mean ± SD). ( n = 9 NC and 175 BA subjects). The p -values were shown in plots and were determined using one-way ANOVA with Tukey’s test ( b – f ) and two-tailed Welch’s t -test ( i ).

Article Snippet: To inhibit TGF-β signaling, 1 mg/kg of A8301 sodium (MedChemExpress, HY-10432A) in 20 μL of DPBS was injected intraperitoneally and daily beginning 6 hours after RRV inoculation.

Techniques: In Vitro, Inhibition, Immunostaining, Staining, Immunofluorescence, Two Tailed Test

Journal: Nature Communications

Article Title: Cellular crosstalk mediated by TGF-β drives epithelial-mesenchymal transition in patient-derived multi-compartment biliary organoids

doi: 10.1038/s41467-025-61442-5

Figure Lengend Snippet: a RRV-induced BA mouse model with biliary injury (biliary-injury model). Newborn were injected intraperitoneally (IP) with PBS or RRV followed by daily PBS or A8301. Created in BioRender. Ayabe, H. (2025) https://BioRender.com/b5m1rv9 . b – e Jaundice and survival rates ( n = 9 control [PBS PBS]/23 BA [RRV PBS]/12 BA-treated [RRV A8301] mice), and total bilirubin and plasma ALT (mean ± SD; n = 6 control/6 BA/5 BA-treated mice) in biliary injury model. f H&E-stained EHBD and liver from day 12-14 biliary-injury model. g Immunostaining of VIM (green), ECAD or CK19 (red) and Nucleus (blue; DAPI) in day-4 and day-7 biliary-injury model. VIM and ECAD or CK19 colocalization in control/BA/BA-treated mice were quantified (ECAD: n = 32/32/41 fields examined over 3 mice [day-4], n = 48/69/60 fields examined over 4 mice [day-7], CK19: n = 24/23/31 fields examined over 3 mice [day-4], n = 49/66/60 fields examined over 4 mice [day-7]) (mean ± SD). h RRV-induced BA mouse model with liver fibrosis (fibrosis model). 3-day old mice were injected IP with PBS or RRV followed by daily PBS or A8301. Created in BioRender. Ayabe, H. (2025) https://BioRender.com/6fpm0xa . i – l Jaundice and survival rates ( n = 24 BA/13 BA-treated mice), total bilirubin and plasma ALT (mean ± SD; n = 6 control/6 BA/5 BA-treated mice) in fibrosis model. i ‡ p < 0.01. Source Data file contains exact p -values. m Representative images of H&E-stained liver and EHBD ( n = 6 control/8 BA/6 BA-treated mice) and Sirius Red-stained liver ( n = 6 mice in each group). Experiments were repeated with similar results. n Tissue expression of TGF-β in human BA. Immunostaining of CK19 (C; red), TGF-β1 (T; white), VIM (V; green) and DAPI (N; blue; nucleus) in a liver biopsy from BA patient. Graphs depict colocalization of CK19 and TGF-β1 or VIM (mean ± SD; n = 24 fields from 3 subjects). The p -values were shown in plots and were determined using two-tailed Fisher’s exact test ( i ) with Bonferroni correction ( b ), Log-rank test ( j ) with Bonferroni correction ( c ), one-way ANOVA with Tukey’s test ( d , e , g , k , l ) and two-tailed Welch’s t -test ( n ).

Article Snippet: To inhibit TGF-β signaling, 1 mg/kg of A8301 sodium (MedChemExpress, HY-10432A) in 20 μL of DPBS was injected intraperitoneally and daily beginning 6 hours after RRV inoculation.

Techniques: Injection, Control, Clinical Proteomics, Staining, Immunostaining, Expressing, Two Tailed Test