Journal: Cell Death & Disease

Article Title: Generation of proliferative hESC-derived grape-clustered hepatocyte organoids with multipolar architecture as regenerative counterpart via synergy of YAP and IGF2 pathways

doi: 10.1038/s41419-026-08635-y

Figure Lengend Snippet: A KEGG pathway analysis of G-heporgs and S-heporgs. B Heatmap showing ligands and receptors in G-heporgs and S-heporgs. C Matched signaling of IGF2 and IGF1R using STRING database. D Expressions of IGF2 and IGF1R were analyzed by RT-qPCR in different organoids. n = 4 biologically independent experiments. E IGF2 secretion from culture medium at different days (day 1 was the first day that observed the generation of G-heporgs). n = 4 biologically independent experiments. F Immunostaining for IGF1R and Ki67. Nuclei were stained with DAPI. Scale bar = 25 μm. G Experimental strategy to evaluate the effect of exogenous IGF2 and LY294002 (PI3K-AKT inhibitor) on G-heporgs. H Representative morphologies of G-heporgs after the treatment with or without IGF2 and LY294002 at different days. Scale bar = 100 μm. I Quantification of the formation efficiency ( n = 4 biologically independent experiments) and diameter ( n = 40 organoids) of G-heporgs at day 14 after different treatments. J Immunostaining of ASGPR, HNF4α, ALB, Ecad and Ki67 for G-heporgs treated with IGF2. Nuclei were stained with DAPI. Scale bar = 50 μm. K Western blot assessment for the expression of p-AKT(Thr308), AKT, p-GSK3β(Ser9) and GSK3β in G-heporgs after different treatments. L Schematic illustration of cell-cell interactions between G-heporgs and S-heporgs. Results were presented as mean ± SD. Statistical significance was determined using one‑way ANOVA followed by Tukey post‑test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The concentrations of IGF2, IL32, and CCL20 were quantified using a Human IGF2 ELISA kit (Jingmei Biotechnology, JM-0862H2), a Human IL32 ELISA kit (SAB), and a Human CCL20 ELISA kit (Jingmei Biotechnology, JM-5919H2), respectively, according to the manufacturers’ instructions.

Techniques: Quantitative RT-PCR, Immunostaining, Staining, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Generation of proliferative hESC-derived grape-clustered hepatocyte organoids with multipolar architecture as regenerative counterpart via synergy of YAP and IGF2 pathways

doi: 10.1038/s41419-026-08635-y

Figure Lengend Snippet: A Representative images of G-heporgs after the treatment with YAP agonist (GA017) or inhibitor (Verteporfin) in optimized heporg medium (OHM). B Relative cell number ( n = 4 biologically independent experiments) and organoid diameter ( n = 30 organoids) of G-heporgs at day 5 after the treatment. C Representative morphologies of G-heporgs after the treatment with GA017 at different days. Scale bar = 100 μm. D Immunostaining of Factin and YAP. Nuclei were stained with DAPI. Scale bar = 50 μm. Yellow arrows indicate YAP with nuclear localization, red arrows indicate YAP without nuclear localization. E The quantification of YAP nuclear localization. n = 4 biologically independent experiments. F H&E staining for G-heporgs treated with or without GA017, and the quantification of the percentage of cysts in organoids. Scale bar = 100 μm. n = 4 biologically independent experiments. G Heatmap of G-heporgs treated with or without GA017 for genes related to proliferation, mature and YAP. H GSEA analysis. I The relative fold increment of G-heporgs in the EM with GA017 or IGF2. Each dot represents a passage event. n = 4 biologically independent experiments. Results were presented as mean ± SD. Statistical significance was determined using two‑way repeated measures ANOVA followed by Tukey’s multiple comparisons test, one‑way ANOVA followed by Tukey post‑test and unpaired two-tailed Student’s t-test. *** p < 0.001.

Article Snippet: The concentrations of IGF2, IL32, and CCL20 were quantified using a Human IGF2 ELISA kit (Jingmei Biotechnology, JM-0862H2), a Human IL32 ELISA kit (SAB), and a Human CCL20 ELISA kit (Jingmei Biotechnology, JM-5919H2), respectively, according to the manufacturers’ instructions.

Techniques: Immunostaining, Staining, Two Tailed Test