Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a Overview of the filtering strategy. b The screened miRNA (mir-24-3p)–mRNA interaction network. c The expression of miR-24-3p in each COCA subtype and normal samples. d S100A8 expression in each COCA subtype and normal samples. e Schematic diagram of the CP mouse model construction, single-cell data processing, and estimation of exosome secretion activity using the sEVtrans algorithm. f Left: UMAP plot of all cell types and identified exosomes (sEVs) in the single-cell sequencing data. Right: proportion of exosomes from the CP group and NC group. g UMAP plot showing the exosome secretion activity index (ESAI) value distributions among different cells. h Violin plot showing the ESAI distributions in different cell types. **** P < 0.0001. i Violin plot showing the expression of S100A8 in different cell types. j Representative images of Ly6G, F4/80 and S100A8 immunofluorescence in the mouse CP model. k Schematic diagram of the CP mouse model constructed with intraperitoneal injection of a neutrophil depletion antibody (anti-Ly6g) compared with that generated by isotype injection. l Representative flow cytometry image after establishment of the CP model compared with that after mock establishment via isotype injection. m Neutrophil counts in CD45 + blood cells in CP mice after model establishment compared with mock establishment. n mRNA expression of exosomal S100A8 in the CP mice after model establishment compared with mock establishment.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Expressing, Activity Assay, Sequencing, Immunofluorescence, Construct, Injection, Generated, Flow Cytometry

Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a Exosomes loaded with PKH26 dye can be taken up by HL-60 neutrophils, including the exosome-supplemented group and the empty control group. b S100A8 mRNA expression in HL-60 cells after transfection with miR-24-3p or S100A8. n = 3 per group. c Protein expression of S100A8 in HL-60 cells after transfection with miR-24-3p mimic or inhibitor. n = 3 per group. d Design of the luciferase reporter vector with sequencing alignment of miR-24-3p with the 3’-UTR of the S100A8 gene. e Luciferase activity of the human S100A8 3’-UTR in HEK293T cells. Cells were transfected with empty plasmid (NC), WT, or mutant 3’-UTR (Mut) plasmids together with miR-24-3p mimics or inhibitors. n = 3 per group. f Schematic diagram of the neutrophil and pancreatic stellate cell co-culture system. Relative protein levels of collagen-1 in pancreatic stellar cells after co-culture with mir-24-3p mimic ( g ), inhibitor ( h ), or paquinimod ( i ). n = 3 per group. j The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8. n = 3 per group. k Schematic diagram of paquinimod administration in the CP mouse model. l Representative HE and Masson sections from CP model mice with or without paquinimod injection (left) and the pathological scores of the two groups. n = 5 per group. m The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8 (1 µg/mL) and transfection with CD36, TLR4, RAGE or NC siRNA. n = 3 per group. n Representative fluorescence images of ROS in pancreatic stellate cells from the control group, S100A8 (1 µg/mL) group, S100A8 (1 µg/mL) + 5 mM NAC group, and S100A8 (1 µg/mL) + 10 mM NAC group. o The protein levels of collagen-1 and FN in pancreatic stellate cells after exogenous supplementation with S100A8 (1 µg/mL) or NAC (5 mM or 10 mM). n = 3 per group.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Control, Expressing, Transfection, Luciferase, Plasmid Preparation, Sequencing, Activity Assay, Mutagenesis, Co-Culture Assay, Injection, Fluorescence

Journal: Cell Discovery

Article Title: Integrated transcriptome profiling of plasma exosomes reveals molecular stratification of exocrine and endocrine disorders and S100A8-mediated cell interactions in chronic pancreatitis

doi: 10.1038/s41421-025-00832-x

Figure Lengend Snippet: a mRNA expression of IL-1B, IL6, and TNF in RAW264.7 cells after exogenous supplementation with S100A8 (1 µg/mL) and transfection with or without TLR4 siRNA. n = 3 per group. b Schematic diagram of the macrophage and pancreatic stellate cell co-culture system. c Relative protein levels of BCL-2 and Bax in MIN6 cells after culture with RAW264.7 cells, TLR4-knockdown RAW264.7 cells, and/or exogenous supplementation with S100A8 (1 µg/mL). n = 3 per group. d Representative immunofluorescence image and TUNEL-positive cell counts of MIN6 cells after co-culture with RAW264.7 cells, TLR4-knockdown RAW264.7 cells, and/or exogenous supplementation with S100A8 (1 µg/mL). n = 5 per group.

Article Snippet: Different level of S100A8 protein (HY- P70531 , MCE company, USA) was used to stimulate the HPSC cells.

Techniques: Expressing, Transfection, Co-Culture Assay, Knockdown, Immunofluorescence, TUNEL Assay