Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Schematic workflow of the single cell type (SiCT) and sequential cell type (SeCT) in vitro models. A , workflow of the single cell type (SiCT) in vitro model. (i) Fasciola hepatica metacercariae were in vitro excysted, and the resulting (ii) FhNEJ parasites were cocultured with mesothelial cells (MCs) and hepatic stellate cells (HSCs) for 3 h at 37 °C in 5% CO 2 . (iii) after the coculture period, the parasite and cells were subjected to protein extraction separately. (iv) cell and parasite controls included for the in vitro model. B , workflow of the sequential cell type (SeCT) in vitro model. (i) first period of coculture for 3 h at 37 °C in a 5% CO 2 atmosphere and (ii) second period of coculture for 3 h at 37 °C in a 5% CO 2 atmosphere, with different combinations of parasites and host cells, as follows: Condition 1 (C1), FhNEJ incubated with SIECs then transferred to MC; Condition 2 (C2), FhNEJ incubated with SIECs then transferred to a cell-free plate; Condition 3 (C3), FhNEJ incubated alone then transferred to MC; Condition 4 (C4), FhNEJ incubated without host cells for 6 h; Condition 5 (C5), SIEC incubated alone for 3 h; Condition 6 (C6), MC incubated alone for 3 h. All conditions were performed in triplicate. SIEC, small intestinal epithelial cell.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: Single Cell, In Vitro, Protein Extraction, Incubation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Volcano plots illustrating differentially expressed proteins (DEPs) identified through proteomic analysis of the single cell type (SiCT) in vitro model. A , cytosolic and membrane proteins in mesothelial cells (MCs) incubated with parasites compared to MCs incubated alone. B , cytosolic and membrane proteins in hepatic stellate cells (HSCs) incubated with parasites compared to HSCs incubated alone. C , somatic and tegumental proteins in FhNEJ incubated with MCs compared to parasites incubated alone. D , somatic and tegumental proteins in FhNEJ incubated with HSCs compared to parasites incubated alone. Proteins showing upregulation, indicating increased expression levels, are represented by red dots , while those showing downregulation, indicating decreased expression levels, are represented by blue dots . The number of DEPs is indicated for each comparison. The gray dashed line denotes the statistical significance threshold (q value < 0.05).

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: Single Cell, In Vitro, Membrane, Incubation, Expressing, Comparison

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Network visualization of enriched biological processes (GO terms) across host cell and FhNEJ fractions in the single cell type (SiCT) in vitro model. A , cytosolic and membrane GO terms in mesothelial cells (MCs) incubated with parasites compared to MCs incubated alone. B , membrane GO terms in hepatic stellate cells (HSCs) incubated with parasites compared to HSCs incubated alone. C , somatic GO terms in FhNEJ incubated with MCs compared to FhNEJ incubated alone. D , somatic GO terms in FhNEJ incubated with HSCs compared to FhNEJ incubated alone. “Value” represents the node score of the GO term in its enrichment, visualized by the node color (from yellow : lower significance to blue : higher significance). “Weight” indicates the relative importance of the GO term, represented by arrow thickness . “Dispensability” reflects term redundancy, where lower values indicate more representative terms within the network, with node size inversely related to dispensability.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: Single Cell, In Vitro, Membrane, Incubation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Differentially expressed proteins (DEPs) in host cells and parasites under varying experimental conditions during sequential cell type (SeCT) in vitro model. A , cytosolic and membrane proteins in intestinal epithelial cells (SIECs) incubated with parasites (C1) versus control not incubated with parasite (C5); B , cytosolic and membrane proteins in mesothelial cells (MCs) incubated with FhNEJ that had been previously incubated with SIECs (C1) compared to MCs incubated with FhNEJ that had been previously maintained alone for 3 h (C3); ( C ) cytosolic and membrane proteins in MCs incubated with parasites from C1 compared to the same cells incubated in the absence of FhNEJ (C6); ( D ) somatic and tegumental proteins in parasites from C1 compared to FhNEJ incubated only with SIECs (C2); ( E ) somatic and tegumental proteins in parasites from C1 compared to FhNEJ incubated only with MC (C3); ( F ) somatic and tegumental proteins in parasites from C1 compared to FhNEJ without host cell incubation (C4). In all panels, proteins showing significant upregulation are indicated with red dots , while significantly downregulated proteins are shown in blue . The number of DEPs identified in each comparison is indicated. The gray dashed line marks the significance threshold (q value < 0.05). The top five upregulated and downregulated proteins associated with each volcano plot are annotated with their Log2 fold changes and expression values across time points, visualized using a red -to- blue gradient scale. Panel E is labeled as “Not detected”, reflecting that no proteins passed the statistical threshold for differential expression in this comparison.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: In Vitro, Membrane, Incubation, Control, Comparison, Expressing, Labeling, Quantitative Proteomics

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Gene Set Enrichment Analysis (GSEA) across cell fractions in the sequential SeCT in vitro model. A , cytosolic and membrane proteins in small intestinal epithelial cells (SIECs) incubated with parasites (C1) versus control not incubated with parasite (C5); B , cytosolic proteins in mesothelial cells (MCs) incubated with FhNEJ that had been previously incubated with SIECs (C1) compared to MC incubated with FhNEJ that had been previously maintained alone for 3 h (C3) or compared to the same cells incubated in the absence of FhNEJ (C6); ( C ) membrane MC proteins comparing C1 versus C3 and C1 versus C6, respectively. Venn diagram showing the overlap of enriched GO terms in the cytosolic fraction and the membrane fraction of MC proteins under the different experimental conditions. Gene Ontology (GO) terms are classified into Biological Process (BP), Cellular Component (CC), and Molecular Function (MF). “% Seq” represents the percentage of proteins associated with each GO term in the dataset. “Size” refers to the number of proteins included in each enriched GO term. “FDR ( p value)” refers to false discovery rate, visualized with a color gradient ( yellow : low significance; blue : high significance). SeCT, sequential cell type; FDR, false discovery rate.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: In Vitro, Membrane, Incubation, Control

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Gene Set Enrichment Analysis (GSEA) across parasite experimental conditions in the sequential cell type (SeCT) in vitro model. A , somatic protein fraction of FhNEJ sequentially incubated with small intestinal cells (SIECs) and mesothelial cells (MCs) (C1) compared to FhNEJ incubated only with SIECs (C2) or compared to FhNEJ incubated only with MCs (C3); (B) tegumental protein fraction comparing C1 versus C2 and C4, respectively. Venn diagram showing the overlap of enriched GO terms in the somatic fraction and the tegumental fraction of mesothelial cells (MCs) proteins under the different experimental conditions. Gene Ontology (GO) terms are classified into Biological Process (BP), Cellular Component (CC), and Molecular Function (MF). “% Seq” represents the percentage of proteins associated with each GO term in the dataset. “Size” refers to the number of proteins included in each enriched GO term. “FDR ( p value)” refers to false discovery rate, visualized with a color gradient ( yellow : low significance; blue : high significance). FDR, false discovery rate.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: In Vitro, Incubation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Heatmaps illustrating the cell proteomic expression profiles of focal adhesion KEGG pathway in the sequential cell type SeCT in vitro model. A , proteomic profile in intestinal epithelial cells (SIECs) incubated with FhNEJ (C1) and SIECs incubated in the absence of FhNEJ (C5), respective squares. B , proteomic profile in mesothelial cells (MCs) incubated with FhNEJ that had been previously incubated with SIEC (C1), MCs incubated with FhNEJ that had been previously maintained alone for 3 h (C3) and MCs incubated alone (C6), respective squares. Each square highlights the specific experimental condition represented in the heatmap , enabling direct visual comparison between groups. Expression levels are shown using a gradient color scale ranging from blue to red , representing low to high abundance, respectively. Membrane-associated proteins are displayed in the membrane compartment of the pathway diagram, while cytosolic proteins appear in the cytosolic compartment. Protein identifiers correspond to KEGG pathway annotations. KEGG, Kyoto Encyclopedia of Genes and Genomes; SeCT, sequential cell type.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: Expressing, In Vitro, Incubation, Comparison, Membrane

Journal: Molecular & Cellular Proteomics : MCP

Article Title: In vitro Host-Parasite Models Combined With Proteomics Reveal Cell Type-Specific Responses of Early Migrating Fasciola hepatica Juveniles

doi: 10.1016/j.mcpro.2026.101543

Figure Lengend Snippet: Heatmap depicting the comprehensive manual annotation of the endopeptidase protein family, and their inhibitors found in the in vitro model for FhNEJ in the sequential cell type (SeCT) in vitro model. The plot displays the proteomic values of each annotated protein, organized by their respective families. Proteases are categorized as cathepsin and legumain in the upper block, while the protease inhibitors are categorized as cysteine peptidases inhibitors (cystatin and stefin), kunitz and serpin in the lower block. The first four squares depict FhNEJ sequentially incubated with small intestinal epithelial cells (SIECs) and mesothelial cells (MCs) of C1, FhNEJ incubated only with SIECs (C2), FhNEJ incubated only with MCs (C3), and FhNEJ without host cell incubation (C4) for somatic proteins, while the subsequent four squares illustrate the same conditions for tegumental proteins. Expression levels are represented visually with a gradient color scale that ranges from blue to red , indicating low to high expression, respectively.

Article Snippet: Cells were cultured in their specific media: Complete Epithelial Cell Medium (Cell Biologics, #M6621) for SIECs, Mouse Mesothelial Cell Culture Complete Medium (Celprogen, #M66233–01S) for MCs, and PriGrow III Medium (ABM, #TM003) supplemented with 10% non–heat-inactivated fetal bovine serum and 1% penicillin-streptomycin for HSCs, at 37 °C with 5% CO 2 in flasks precoated with 0.2% extracellular matrix (ECM) (Cell Biologics, #6950).

Techniques: In Vitro, Blocking Assay, Incubation, Expressing