Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Single cell sequencing and immunohistochemistry validate key molecular markers for BA. (A) Representative immunohistochemistry images validating the protein expression of mitophagy-related genes ( IGF2BP3 , NEDD4L , and ALDH2 ) in liver tissues from BA patients and controls (scale bar =100 µm). Ctrl images are from non-tumor liver tissue adjacent to hepatoblastoma specimens. Positive staining appears as brown discoloration and infiltrating immune cells were indicated by red arrows. (B) Differential analysis of single-cell transcription. (a) Single-cell’s cell classification landscape in all samples. (b) The single-cell expression differences of IGF2BP3, NEDD4L, PPIB, and ALDH2 between BA and Ctrl groups. BA, biliary atresia; Ctrl, control; FC, fold change.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Single Cell, Sequencing, Immunohistochemistry, Expressing, Staining, Control

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: The interaction between drugs and IGF2BP3 and NEDD4L proteins. (A-C) The interaction between cyclosporine and IGF2BP3 protein. (D-F) The interaction between cyclosporine and NEDD4L protein. Blue represents drug molecules, with red indicating oxygen atoms within the molecules. Green represents amino acid residues of the protein interacting with the drug molecules. The yellow background indicates the average interaction energy between the protein and drugs (average of 10 simulations).

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques:

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Potential schematic representation of the BA-CIMDGM genes in the pathogenesis of BA. Upregulated IGF2BP3 in T cells and NEDD4L in macrophages leads to impaired mitophagy and promotes immune activation and cytokines released. And the downregulation of ALDH2 in hepatocytes compromises the mitochondrial antioxidant defense and excessive ROS production, synergistically driving biliary epithelial cell injury and progressive liver fibrosis. BA, biliary atresia; BA-CIMDGM, Biliary Atresia Cell Infiltration Mitophagy Diagnostic Gene Model; ROS, reactive oxygen species.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Activation Assay, Diagnostic Assay

Journal: Nature Communications

Article Title: Human iPS-derived pre-epicardial cells direct cardiomyocyte aggregation expansion and organization in vitro

doi: 10.1038/s41467-021-24921-z

Figure Lengend Snippet: a Schematics timeline and protocol of ventricular-like CM differentiation from lateral plate mesoderm (LPM, Top), and its cardiac gene expression at day 30 (Bottom) from three independent differentiations. Data presented as mean ± SEM. Unpaired ttest with Welch’s correction was used for statistical comparison between groups. b Representative immunophenotypic characterization of ventricular-CM at day 30 from three independent differentiation. BMS = retinoid-x-receptor antagonist BMS189453. c Representative immunofluorescence images of Sarcomeric Actinin (Red) EdU + (green) ventricular CMs after co-culturing with PECs from three independent experiments. d Representative flow cytometric quantification of cTnT + Edu + ventricular CM in PEC co-culture after 6 days. The percentage in red was calculated by taking the cell count in Q2 and dividing by 100% cTnT + cells, or total events in Q1 and Q2 only (red box). Data presented in mean ± SEM ( n = 3). Two-tailed student T test was used. e Percentage of cTnT + EdU + VM in response to Linsitinib. Data presented in mean ± SEM ( n = 3 independent experiments) versus 0 µM Linsitinib. One-way ANOVA with Dunnett post hoc test was used. f Dose-response examination of IGF2 on CM proliferation. Data presented in mean ± SEM ( n = 3 independent experiments). One-way ANOVA with Dunnett post hoc test was used. g ELISA analysis of IGF2 in conditioned medium 2 and 6 days after co-culturing with PECs. Data presented in mean ± SEM ( n = 3 independent experiments). One-way ANOVA with Dunnett post hoc test was used. **** p value < 0.0001 versus CM only. h The expression of IGF2 RNA in PECs in response to increasing CM number. Data presented in mean ± SEM ( n = 3 independent experiments). One-way ANOVA with Dunnett post hoc test was used. **** p value < 0.0001 versus no CMs. i IGF2 expression in PECs in CM co-culture in transwell with or without RARα/γ antagonist BMS-189453 (BMS). Data presented in mean ± SEM ( n = 3 independent experiments). One-way ANOVA with Dunnett post hoc test was used. **** p < 0.0001 versus PECs-CM co-culture without BMS.

Article Snippet: IGF2 protein were quantified using Human IGF-II/IGF2 Quantikine ELISA kit DG200(R&D Systems, Minneapolis, MN).

Techniques: Expressing, Comparison, Immunofluorescence, Co-Culture Assay, Cell Counting, Two Tailed Test, Enzyme-linked Immunosorbent Assay