Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: IGF2BP3 is increased in type 2 disease. A. Left: normalized expression levels of IGF2BP3 mRNA in severe asthma (SA) vs healthy control (HC) primary BECs at total and polyribosome-bound levels; centre: IGF2BP3 normalized microarray expression data comparing airway epithelial cells from healthy non-atopic (HNA) and atopic asthma (AA) children; right IGF2BP3 normalized microarray expression data comparing nasal lavage samples from children during Picornavirus-induced exacerbation (EX) vs 7-14 days post-exacerbation (post-EX). B. Normalized count levels comparing type 2 high vs type 2 low nasal brushings for genes that showed IGF2BP3-dependent IL-13-modulation in our Frac-seq data . C: Heatmap showing the normalized expression values of the common mRNA signature between differentially expressed genes in the GALA II cohort and our Frac-seq IGF2BP3 and IL-13-dependent DEGs. D. Model for IGF2BP3 effects on type 2 responses. Decreasing IGF2BP3 levels leads to increased stability of IL13RA1 and IL4R mRNA and increased surface expression of IL4R and IL13Rα1, leading to increased STAT6 phosphorylation and increased IL-13 (and possibly IL-4) transcriptional responses, contributing to an overall increased type 2 response. In disease, IGF2BP3 is upregulated potentially to supress excessive T2 responses.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Expressing, Control, Microarray, Phospho-proteomics

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: Decreasing IGF2BP3 levels increases IL-13/IL-4-driven transcriptional responses. RT-qPCR results of primary BECs transfected with siRNAs against IGF2BP3 (siIGF2BP3) or scrambled control (siControl) and stimulated with IL-13 or IL-4. A: RT-qPCR results for IGF2BP3 knock down and the major signalling mediators of IL-13 and IL-4. B: RT-qPCR results for chemokines. C: RT-qPCR results for pro-inflammatory IL6 and IL8 mRNAs. Data (n=3) depicted as mean ±SEM. One-tailed ratio t-tests. * P ≤ 0.05; ** P ≤ 0.01.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Quantitative RT-PCR, Transfection, Control, Knockdown, One-tailed Test

Journal: eBioMedicine

Article Title: Polymeric immunoglobulin receptor promotes Th2 immune response in the liver by increasing cholangiocytes derived IL-33: a diagnostic and therapeutic biomarker of biliary atresia

doi: 10.1016/j.ebiom.2024.105344

Figure Lengend Snippet: (A) Immunostaining for PIGR (Red) and CK19 (Green) in liver and EBD sections from the BA and control patients. Original magnification, × 400; Scale bars, 50 μm (B) Quantification of the fluorescence intensity of PIGR signals in the liver and EBD sections from the BA and control patients (n = 6; ∗∗, P < 0.01; Wilcoxon rank sum test); expression of PIGR mRNA in the liver and EBD of BA and control patients (n = 12; ∗∗∗, P < 0.001; Wilcoxon rank sum test) (C) Schematic illustration depicting the three experimental mouse models employed in this study (created with BioRender.com ), and the representative images of each mouse model (D) Masson staining of liver sections from the experimental mouse models (E) Expression of PIGR in the liver of experimental mouse models detected by immunostaining, RT-qPCR and immunoblotting analyses (n = 6; ns, P > 0.05; ∗, P < 0.05; ∗∗∗, P < 0.001; Wilcoxon rank sum test) (F) Expression of PIGR in cultured BECs upon stimulations of different cytokines, detected by RT-qPCR and immunoblotting analyses, and immunostaining (n = 3; ns, P > 0.05; ∗, P < 0.05; ∗∗∗, P < 0.001; two-tailed t-test) (G) Expression of PIGR in cultured BECs upon stimulations of different cytokines, with or without the pretreatment of a NF-κB inhibitor PDTC; detected by RT-qPCR and immunoblotting analyses, and immunostaining (n = 3; ns, P > 0.05; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; two-tailed t-test).

Article Snippet: Human intrahepatic biliary epithelial cells (BECs) (#5100, ScienCell) were cultured in RPMI 1640 medium (#PM150110, Pricella Life Science &Technology Co., Ltd) supplemented with 10% fetal bovine serum (#164210, Pricella).

Techniques: Immunostaining, Control, Fluorescence, Expressing, Staining, Quantitative RT-PCR, Western Blot, Cell Culture, Two Tailed Test

Journal: eBioMedicine

Article Title: Polymeric immunoglobulin receptor promotes Th2 immune response in the liver by increasing cholangiocytes derived IL-33: a diagnostic and therapeutic biomarker of biliary atresia

doi: 10.1016/j.ebiom.2024.105344

Figure Lengend Snippet: (A) Heatmap and volcano plot of the differential expressed genes between vector control BECs (vector) and PIGR overexpressed BECs (Over) (B) Detection of IL-33 expression in PIGR-silenced, PIGR-overexpressed, and corresponding control BECs, using immunostaining (original magnification, × 400; scale bars, 50 μm) and RT-qPCR analysis (n = 3; ∗, P < 0.05; ∗∗, P < 0.01; two-tailed t-test); quantification of supernatant levels of IL-33 of the PIGR-silenced, PIGR-overexpressed, and corresponding control BECs, using enzyme-linked immunosorbent assay (n = 3; ∗, P < 0.05; ∗∗, P < 0.01; two-tailed t-test) (C) Immunofluorescence staining of CK19 (green), PIGR (red), and IL-33 (pink) in the liver sections of PIGR-silenced, PIGR-overexpressed, and corresponding control mice on day 14 (original magnification, × 400; scale bars, 100 μm) (D) Detection of PIGR mRNA and protein levels in the liver tissue of mice on day 14 (n = 6; ∗, P < 0.05; ∗∗, P < 0.01; Wilcoxon rank sum test) (E) Detection of IL-4, IL-5, IL-10, and IL-13 concentrations in the liver homogenate of PIGR-silenced mice (with or without injection of exogenous IL-33) using enzyme-linked immunosorbent assay (n = 6; ∗∗∗, P < 0.001; Wilcoxon rank sum test); FCM analyses of Th2 cell proportion in isolated hepatic lymphocytes of mice on day 14, the results represent one of five independent experiments (n = 5; ∗∗, P < 0.01; Wilcoxon rank sum test) (F) Correlation analyses of concentrations of PIGR and four cytokines in the liver homogenate of BA patients (n = 60); r, Pearson correlation coefficient; P, P-value analyzed with two-tailed t-test.

Article Snippet: Human intrahepatic biliary epithelial cells (BECs) (#5100, ScienCell) were cultured in RPMI 1640 medium (#PM150110, Pricella Life Science &Technology Co., Ltd) supplemented with 10% fetal bovine serum (#164210, Pricella).

Techniques: Plasmid Preparation, Control, Expressing, Immunostaining, Quantitative RT-PCR, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Injection, Isolation

Journal: eBioMedicine

Article Title: Polymeric immunoglobulin receptor promotes Th2 immune response in the liver by increasing cholangiocytes derived IL-33: a diagnostic and therapeutic biomarker of biliary atresia

doi: 10.1016/j.ebiom.2024.105344

Figure Lengend Snippet: (A) Validation of the efficiencies of silencing and overexpressing PIGR in cultured BECs by detecting PIGR expression using RT-qPCR and immunoblotting analyses (n = 3; ∗∗∗, P < 0.001; two-tailed t-test), and immunostaining (original magnification, × 400; scale bars, 50 μm) (B) Proliferation of PIGR-silenced and PIGR-overexpressed BECs from 0 to 72 h post-transfection (n = 3; ns, P > 0.05; ∗, P < 0.05; ∗∗, P < 0.01; two-tailed t-test). Data are presented as mean ± SEM of three independent experiments (C) Apoptosis of PIGR-silenced and PIGR-overexpressed BECs detected by FCM after staining with Annexin-V-FITC and PI (n = 3; ns, ∗∗, P < 0.01; ∗∗∗, P < 0.001; two-tailed t-test). Data presented are as mean ± SEM of three independent experiments (D) Detection of two EMT markers (α-SMA and E-cadherin) in PIGR-silenced and PIGR-overexpressed BECs, using RT-qPCR and immunoblotting analyses (n = 3; ∗, P < 0.05; ∗∗, P < 0.01; two-tailed t-test), and immunostaining (original magnification, × 200; scale bars, 50 μm).

Article Snippet: Human intrahepatic biliary epithelial cells (BECs) (#5100, ScienCell) were cultured in RPMI 1640 medium (#PM150110, Pricella Life Science &Technology Co., Ltd) supplemented with 10% fetal bovine serum (#164210, Pricella).

Techniques: Biomarker Discovery, Cell Culture, Expressing, Quantitative RT-PCR, Western Blot, Two Tailed Test, Immunostaining, Transfection, Staining