Journal: bioRxiv

Article Title: TL1A-activated T cells remodel the rectal mucosa in Crohn’s disease patients with perianal fistulizing disease

doi: 10.1101/2025.06.26.657455

Figure Lengend Snippet: A: Violin plot for LTB expression in the T-cell compartment. The adjusted p-values obtained by DEG analysis are indicated for each comparison. B: ELISA for TFPI2 in the supernatants of primary intestinal fibroblasts stimulated with LTα 1 β 2 (100ng/ml) for 24 hours compared to the unstimulated condition. Paired T test, nonparametric, n=10, ** = p < 0.01. C: Violin plot for LTB and TNFRSF25 expression in the CD4+ T cell compartment, comparing CDun and CDinf patients. The adjusted p-values generated by DEG analysis are indicated. D: Violin plot for LTB and TNFRSF25 expression in the CD4+ T cell compartment, comparing CDinf and CD+PFDinf patients. The adjusted p-values generated by DEG analysis are indicated. E: On the left, Uniform Manifold Approximation and Projection (UMAP) plots of the whole object showing the expression of TNFSF15 . On the right, violin plots indicating the expression of TNFSF15 . F: Graphical representation of the flow cytometry approach to detect LTα 1 β 2 + T cells.

Article Snippet: Supernatants of stimulated fibroblasts and tissue explants were used for cytokine detection using the DuoSet ELISA Kits from R&D for human MMP1 (DY901B) and IL-22 (DY782), and the human TFPI2 ELISA Kit from MyBioSource (MBS2507217) according to the manufacturer’s instructions.

Techniques: Expressing, Comparison, Enzyme-linked Immunosorbent Assay, Generated, Flow Cytometry

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Relationship between the regulation of TFPI2 expression in cancer cells and host cells. The signaling pathway constituents numbered 1 to 21 in the figure are described in sections of the text labeled as (1,2), and so forth. Arrows denote activation, while T-shaped symbols represent inhibition. ACTN4, actinin 4; AP-1, activator protein 1; AP-2α, activator protein 2 alpha; CLIP1, CAP-Gly domain-containing linker protein 1 (CLIP1); COX-2, cyclooxygenase-2; DNMT1, DNA methyltransferase 1; EGF, epidermal growth factor; ERK, extracellular signal-regulated kinase; HGF, hepatocyte growth factor; IL-10, interleukin-10; JNK, c-Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; MBD3, methyl-CpG-binding domain protein 3; MMP, matrix metalloproteinase; MYH9, myosin 9; MITF, melanocyte-induced transcription factor; NF-κB, nuclear factor kappa B; PPARγ, peroxisome proliferator-activated receptor gamma; PSAP, prosaposin; SLUG, snail family transcriptional repressor 2; TAM, tumor-associated macrophages; TFPI2, tissue factor pathway inhibitor 2; TGF-β, transforming growth factor beta; TGFBI, transforming growth factor beta-induced; TIRAP, TIR domain containing adaptor protein; TMPRSS4, transmembrane protease serine 4; uPA, urokinase-type plasminogen activator; VEC, vascular endothelial cells; VEGF, vascular endothelial growth factor; VSMC, vascular smooth muscle cells.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Expressing, Labeling, Activation Assay, Inhibition, Binding Assay

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Molecules and signaling pathways regulated by TFPI2. The left pie chart illustrates molecules and signaling pathways modulated by TFPI2 that are implicated in the regulation of glucose and lipid metabolism, macrophage polarization, inflammatory responses, immune modulation, and cellular proliferation and survival. In contrast, the right pie chart depicts molecules and signaling cascades associated with extracellular matrix remodeling, cytoskeletal integrity, and cellular motility.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Protein-Protein interactions

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Molecular and signaling pathways governing the regulation of TFPI2 expression. The left pie chart depicts factors that promote the upregulation of TFPI2, while the right pie chart illustrates those that facilitate its downregulation. These genes contribute to cancer invasion and metastasis by modulating inflammatory responses, promoting angiogenesis, regulating immune functions, and influencing tumor cell proliferation, survival, and apoptotic pathways.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Protein-Protein interactions, Expressing

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: The role of TFPI2 in pregnancy-induced hypertension, diabetes, and atherosclerosis. TFPI2 orchestrates extracellular matrix (ECM) remodeling, tissue regeneration, angiogenesis, and immune modulation. The preservation or disturbance of cellular homeostasis through inflammatory processes may result in TFPI2 dysregulation, thereby contributing to the onset and progression of diabetes, atherosclerosis, and pregnancy-induced hypertension. Superscript arrows represent the upregulation of each factor or the activation of pathological or functional processes, whereas subscript arrows denote downregulation or functional inhibition. DNMT1, DNA methyltransferase 1; ERK, extracellular signal-regulated kinase; EZH1, enhancer of zeste 1 polycomb repressive complex 2 subunit; H3K27me3, tri-methylation at lysine 27 of histone H3; IL-1, interleukin-1; MAPK, mitogen-activated protein kinase; MMP, matrix metalloproteinase; MΦ, macrophage; PI3K, phosphatidylinositol-3-kinase; PPARγ, peroxisome proliferator-activated receptor gamma; TFPI2, tissue factor pathway inhibitor 2; TGF-β, transforming growth factor-beta; TNF-α, tumor necrosis factor-alfa; VEC, vascular endothelial cells; VEGFR, vascular endothelial growth factor receptor; VSMC, vascular smooth muscle cells.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Preserving, Activation Assay, Functional Assay, Inhibition, Methylation

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Role of TFPI2 in cancer invasion and metastasis. As illustrated in the upper left panel, TFPI2 overexpression suppresses ECM remodeling and inhibits tumor invasion in vitro. Conversely, the upper right panel shows that TFPI2 downregulation facilitates invasion through MMP-mediated ECM degradation. In vivo, TFPI2 influences not only cancer cells but also diverse components of the TME, including inflammatory and immune cells, endothelial cells, cytokines, ECM constituents, and angiogenic factors, thereby playing a pivotal role in modulating tumor malignancy and metastatic dissemination. Superscript arrows represent the upregulation of each factor or the activation of pathological or functional processes, whereas subscript arrows denote downregulation or functional inhibition.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Over Expression, In Vitro, In Vivo, Activation Assay, Functional Assay, Inhibition

Journal: Experimental & Molecular Medicine

Article Title: Hypothermic oxygenated perfusion inhibits CLIP1-mediated TIRAP ubiquitination via TFPI2 to reduce ischemia‒reperfusion injury of the fatty liver

doi: 10.1038/s12276-024-01350-8

Figure Lengend Snippet: a , b Western blotting and statistical analyses of the TFPI2 protein in liver tissue ( n = 3 per group). c RT‒qPCR was used to detect TFPI2 mRNA levels in liver tissue ( n = 3 per group). d , e IHC staining and statistical analysis of TFPI2 in liver tissue ( n = 5 per group). f Diagram of the SD rat fatty liver model and AAV8 injection. ov-TFPI2, sh-TFPI2, or empty vector AAV8 was injected into SD rats through the tail vein; an MCD diet was started on the second day after the AAV8 injection, and the liver was retrieved on the 15th day for the following experiments. g , h Concentrations of ALT and AST in the perfusate ( n = 5 per group). i , j HE staining and histological necrotic area analysis of liver tissue ( n = 5 per group). k Schematic diagram of the TLR4 and NF-κB inflammatory signaling pathways. i‒o Western blotting and statistical analyses of TFPI2, IL-1β, TNF-α, HMGB1, TLR4, p65, p-p65, lkB, p-lkB, and β-actin proteins in liver tissue ( n = 3 per group). p–s IL-1β, IL-6, IL-10, and TNF-α concentrations in liver tissue as determined by ELISA ( n = 5 per group). The data are presented as the mean ± SD; ns not significant; * P < 0.05; and ** P < 0.01.

Article Snippet: TFPI2-overexpressing (ov-TFPI2), TFPI2-knockdown (sh-TFPI2), and empty vector (NC) of adeno-associated virus serotype 8 (AAV8) were provided by OBiO Technology Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Immunohistochemistry, Injection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay

Journal: Experimental & Molecular Medicine

Article Title: Hypothermic oxygenated perfusion inhibits CLIP1-mediated TIRAP ubiquitination via TFPI2 to reduce ischemia‒reperfusion injury of the fatty liver

doi: 10.1038/s12276-024-01350-8

Figure Lengend Snippet: a Schematic diagram of the identification of proteins that interact with TFPI2 via CoIP combined with LC‒MS. b Liver tissue lysate was incubated with an anti-IgG antibody (IgG) or anti-TFPI2 antibody (IP). Different bands of anti-TFPI2 and anti-IgG were detected in silver-stained SDS‒PAGE gels. c The interaction between TFPI2 and CLIP1 was confirmed by CoIP. d IF analysis of TFPI2 and CLIP1 in HUVECs ( n = 5 per group). e , f Western blotting and statistical analyses of the TFPI2 and CLIP1 proteins in liver tissue ( n = 3 per group). g , h Western blotting and statistical analyses of the CLIP1 protein in the liver tissue of rats infected with ov-TFPI2, sh-TFPI2, or empty vector AAV8 (n = 3 per group). i Schematic diagram of the full-length and deleted domain architecture of TFPI2. The KD1 domain is located in amino acid (aa) sequences 36 to 86, the KD2 domain is located in aa sequences 96 to 149, and the KD3 domain is located in the sequence 158 to 208. Δ means deleted. j , k CoIP analysis of the domains and sites where CLIP1 interacts with TFPI2. Flag-tagged CLIP1, His-tagged TFPI2, KD1, KD2, KD3, and mutant TFPI2 (R24Q) plasmids were transfected into HEK293T cells. The cell protein extract was tested via CoIP with an anti-FLAG primary antibody. The data are presented as the mean ± SD; ns not significant; * P < 0.05; and ** P < 0.01.

Article Snippet: TFPI2-overexpressing (ov-TFPI2), TFPI2-knockdown (sh-TFPI2), and empty vector (NC) of adeno-associated virus serotype 8 (AAV8) were provided by OBiO Technology Co., Ltd. (Shanghai, China).

Techniques: Incubation, Staining, Western Blot, Infection, Plasmid Preparation, Sequencing, Mutagenesis, Transfection

Journal: Experimental & Molecular Medicine

Article Title: Hypothermic oxygenated perfusion inhibits CLIP1-mediated TIRAP ubiquitination via TFPI2 to reduce ischemia‒reperfusion injury of the fatty liver

doi: 10.1038/s12276-024-01350-8

Figure Lengend Snippet: a‒d Western blotting and analysis of TFPI2, CLIP1, IL-1β, TNF-α, HMGB1, p65, p-p65, lkB, p-lkB, and β-actin proteins in HUVECs treated with LPS and transfected with TFPI2, CLIP1, or empty vector. e–g The concentrations of IL-1β, IL-6, and TNF-α in the culture medium of HUVECs treated with LPS and transfected with TFPI2, CLIP1, or empty vector were determined via ELISA. h NF-κB-responsive reporter activity in HEK293T cells transfected with TFPI2 and CLIP1. i and j Apoptosis of HUVECs treated with LPS and transfected with TFPI2, CLIP1, or empty vector detected by flow cytometry. n = 3 per group; the data are presented as the mean ± SD; ns not significant; * P < 0.05; and ** P < 0.01.

Article Snippet: TFPI2-overexpressing (ov-TFPI2), TFPI2-knockdown (sh-TFPI2), and empty vector (NC) of adeno-associated virus serotype 8 (AAV8) were provided by OBiO Technology Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Activity Assay, Flow Cytometry

Journal: Experimental & Molecular Medicine

Article Title: Hypothermic oxygenated perfusion inhibits CLIP1-mediated TIRAP ubiquitination via TFPI2 to reduce ischemia‒reperfusion injury of the fatty liver

doi: 10.1038/s12276-024-01350-8

Figure Lengend Snippet: a , b Western blotting and statistical analyses of the TIRAP protein in the liver tissue of rats infected with ov-TFPI2, sh-TFPI2, or the empty vector AAV8. c , d Western blotting and statistical analyses of the TIRAP and TFPI2 proteins in HUVECs treated with LPS and transfected with lentivirus of sh-TFPI2, plasmid of TFPI2, or empty vector. e , f Western blotting of the TIRAP and TFPI2 proteins in HUVECs transfected with sh-TFPI2 or empty vector and treated with CHX for different times and the degradation curves of the TIRAP protein. g, h Western blotting and statistical analyses of the TIRAP and TFPI2 proteins in HUVECs transfected with sh-TFPI2 or empty vector and treated with MG132 or chloroquine. i Analysis of TFPI2 regulating endogenous TIRAP ubiquitination. Rats were infected with ov-TFPI2, sh-TFPI2, or the empty vector AAV8. Liver tissue protein extracts were tested by CoIP with an anti-TIRAP antibody. j , k Analysis of TFPI2 and CLIP1 regulating exogenous TIRAP ubiquitination. HEK293T cells were transfected with Myc-tagged TIRAP, Flag-tagged CLIP1, HA-tagged Ub, or si-TFPI2. Cell protein extracts were subjected to CoIP with an anti-Myc antibody. l CoIP assays of TIRAP and CLIP1 expression in HEK293T cells with or without TFPI2 overexpression. m IF analysis of CLIP1 and TIRAP in HUVECs with or without TFPI2 overexpression. n = 3 per group; the data are presented as the mean ± SD; ns not significant; * P < 0.05; and ** P < 0.01.

Article Snippet: TFPI2-overexpressing (ov-TFPI2), TFPI2-knockdown (sh-TFPI2), and empty vector (NC) of adeno-associated virus serotype 8 (AAV8) were provided by OBiO Technology Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Infection, Plasmid Preparation, Transfection, Expressing, Over Expression

Journal: Experimental & Molecular Medicine

Article Title: Hypothermic oxygenated perfusion inhibits CLIP1-mediated TIRAP ubiquitination via TFPI2 to reduce ischemia‒reperfusion injury of the fatty liver

doi: 10.1038/s12276-024-01350-8

Figure Lengend Snippet: a Macroscopic view of the human liver; HE staining, ORO staining, and IHC staining of TFPI2 and CLIP1. b–d Western blotting and statistical analyses of the TFPI2 and CLIP1 proteins in human liver tissue. e Analysis of the necrotic area via human liver histology. f Analysis of the ORO-positive area in human liver tissue. g , h Quantitative analysis of IHC staining of TFPI2 and CLIP1 in human liver tissue. n = 5 per group; the data are the mean ± SD; ns, not significant; * P < 0.05; and ** P < 0.01. i Mechanistic scheme. In a rat model of fatty liver, severe steatosis, cold ischemia, and subsequent reperfusion led to the accumulation of ROS in hepatocytes, with the released HMGB1 activating the TLR4/NF-κB inflammatory pathway in ECs. Additionally, R24 of the KD1 domain of TFPI2 directly binds to CLIP1 in ECs, inhibiting the ubiquitination and degradation of TIRAP by CLIP1, thereby activating the TLR4/NF-κB-mediated inflammatory response, resulting in severe IRI. HOPE effectively ameliorates the entire pathological process.

Article Snippet: TFPI2-overexpressing (ov-TFPI2), TFPI2-knockdown (sh-TFPI2), and empty vector (NC) of adeno-associated virus serotype 8 (AAV8) were provided by OBiO Technology Co., Ltd. (Shanghai, China).

Techniques: Staining, Immunohistochemistry, Western Blot