Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet: Combination of CITE-seq, scRNA-seq, snRNA-seq, and spatial analyses enables identification of all hepatic cell types including bona fide cell doublets, related to Figure 1 (A and B) Top DEGs (A) and DEPs (B) for cell types from Figure 1 B. (C) Distinct profiles of cells or nuclei within the UMAP depending on isolation protocols; 71,162 cells from ex vivo digestions, 96,066 cells from in vivo digestions, and 18,666 nuclei. Numbers on plots represent numbers of cells/nuclei per population. (D) Correlation plots showing genes captured within the KC, B cell and neutrophil populations with and without addition of CITE-seq antibodies. (E) Expression of VSIG4, CD206, and ESAM (protein, top) and Vsig4 , Mrc1 , and Esam (mRNA, bottom). (F) UMAP showing clusters of cells when only minimal QC for gene number and % mitochondrial genes is performed; 17,669 cells pooled from 3 samples. Expression of Cd5l, Cd19 , and Kdr by the clusters facilitating identification of cell types per annotation. (G) CITE-seq data from (F) in Flow-Jo showing expression of CD206 and ESAM in total KCs (left) and total B cells (middle). Numbers represent % of entire KC or B cell population. Identified populations were then mapped back onto the original UMAP (right). (H) Expression of CD31, CD26, and CD38 by indicated populations. (I) Heatmaps showing expression of top DEGs between KC1s and LSECs (left), KC2s and KC1s + LSECs (middle) and B cell2s and B cell1s + LSECs (right). (J) 3D reconstruction of murine liver following perfusion with antigen fix to inflate endothelial cells and staining with antibodies against CD31, CD206, and F4/80. (K) UMAP showing clusters generated from Visium analysis of liver tissue (4 samples) and liver capsule (1 sample). (L) Top unbiased genes defining zonation trajectory from portal to central vein in Visium. (M) Expression of Glul and Epcam by confocal microscopy (left), annotation of portal, periportal, mid, and central regions on same tissue section (middle) and overlay of both datasets (right). (N) Identification of cholangiocyte (left) and cDC (right) signatures on zonated Visium spots. (P) Molecular Cartography showing expression of indicated zonated hepatocyte mRNAs in liver tissue. Data are representative of 2 mice. (O) Expression of Itgae (encoding CD103) in the UMAP of the total liver (left) and flow cytometric analysis of total cDC1s for CD103 and MHCII expression in the healthy murine liver (right).

Article Snippet: Anti-Mouse CD26 (REA1196) PE , Miltenyi Biotec , 130-122-775; RRID: AB_2801934.

Techniques: Isolation, Ex Vivo, In Vivo, Expressing, Staining, Generated, Confocal Microscopy

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Mouse CD26 (REA1196) PE , Miltenyi Biotec , 130-122-775; RRID: AB_2801934.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

Journal: Drug Metabolism and Disposition

Article Title: Wuzhi Tablet ( Schisandra Sphenanthera Extract) Protects against Acetaminophen-Induced Hepatotoxicity by Inhibition of CYP-Mediated Bioactivation and Regulation of NRF2-ARE and p53/p21 Pathways

doi: 10.1124/dmd.114.059535

Figure Lengend Snippet: WZ possesses marked inhibitory effects on the activities of P450 enzymes and NAPQI-GSH formation. (A) Western blot analysis of CYP2E1, CYP1A2, CYP3A11, and glyceraldehyde-3-phosphate dehydrogenase levels in livers from control, WZ (700 mg/kg)-treated, APAP-treated, WZ (700 mg/kg)/APAP-treated mice. (B) Densitometric analysis of Western blots (n = 3). (C) Effect of WZ on the activities of CYP2E1, CYP1A2, CYP3A11 enzymes in mouse liver microsomes (n = 5). (D) Effect of WZ on NAPQI-GSH formation in mouse liver microsomes (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001 versus control mice; #P < 0.05, ##P < 0.01 versus APAP-treated mice.

Article Snippet: CYP1A2 and CYP2E1 antibodies were purchased from Boster Biotechnology Co., Ltd. (Wuhang, China).

Techniques: Western Blot, Control

Journal: Stem Cell Research & Therapy

Article Title: CD26 is a senescence marker associated with reduced immunopotency of human adipose tissue-derived multipotent mesenchymal stromal cells

doi: 10.1186/s13287-022-03026-4

Figure Lengend Snippet: Late passage (LP) MSC(AT) have a senescent transcriptome signature. A Gene set enrichment analysis demonstrates increased expression of genes known to be upregulated in senescence (shown in red) and reduced expression of genes downregulated in senescence (shown in blue) in LP-MSC(AT) and shows DPP4 among core enrichment genes; n = 1 pediatric and n = 1 adult, where EP = p6 and LP = p30. B – G Relative expression of selected transcripts from RNAseq analysis tested by quantitative RT-PCR over HPRT housekeeping gene in 9 additional samples. Expression of B CDKN2A (p16), C CDKN1A (p21), D DPP4 ( CD26), E HES1 , F SPP1 and G COL4A1 over the house-keeping gene HPRT . Data presented as mean ± SD, comparisons done with paired Wilcoxon test, * p < 0.05, ** p < 0.01

Article Snippet: The following primary antibodies were used at the dilutions indicated: CD26 (OriGene Technologies Inc., MD, USA; diluted 1:500), β-actin (Chemicon MAB1501; diluted 1: 100,000).

Techniques: Expressing, Quantitative RT-PCR

Journal: Stem Cell Research & Therapy

Article Title: CD26 is a senescence marker associated with reduced immunopotency of human adipose tissue-derived multipotent mesenchymal stromal cells

doi: 10.1186/s13287-022-03026-4

Figure Lengend Snippet: CD26 surface levels and total protein abundance in MSC(AT) increase with replicative senescence and age. A Western Blot of CD26 protein levels in MSC(AT) extracts from early passage (EP) and late passage (LP) pediatric ( n = 2) and adult ( n = 3) samples. Data represent mean + SEM, comparisons done with two-tailed Student’s t -test, * p < 0.05; ** p < 0.01. B Representative immunohistochemistry image of CD26 protein abundance in EP and LP-MSC(AT). C CD26 geometric mean fluorescence intensity (gMFI) and % CD26 high MSCs in EP and LP-MSC(AT) (6 adult and 4 pediatric MSC(AT), at EP = p4.1 ± 0.6 and LP = p21.0 ± 6.0). Data represent mean ± SD, comparisons done with paired Wilcoxon tests, ** p < 0.01. D CD26 gMFI and % CD26 high MSC(AT) in EP-MSC(AT) from pediatric and adult donors (6 pediatric MSC(AT) at p = 4.7 ± 0.5 and 6 adult MSC(AT) p = 3.8 ± 0.4). Data represent mean ± SD, comparisons done with unpaired Mann–Whitney tests, ** p < 0.01

Article Snippet: The following primary antibodies were used at the dilutions indicated: CD26 (OriGene Technologies Inc., MD, USA; diluted 1:500), β-actin (Chemicon MAB1501; diluted 1: 100,000).

Techniques: Western Blot, Two Tailed Test, Immunohistochemistry, Fluorescence, MANN-WHITNEY

Journal: Stem Cell Research & Therapy

Article Title: CD26 is a senescence marker associated with reduced immunopotency of human adipose tissue-derived multipotent mesenchymal stromal cells

doi: 10.1186/s13287-022-03026-4

Figure Lengend Snippet: CD26 high MSC(AT) are less immunosuppressive than CD26 low MSC(AT). A Flow cytometry gating strategy for FACS separation of MSC(AT) based on CD26 surface abundance: CD26 low and CD26 high populations. B Immunopotency assay (i.e., MSC inhibition of proliferating CD4 + T cells). CD3CD28 activated T-cells were co-cultured with either CD26 low or CD26 high MSC(AT) at a 1:16 MSC:PBMC ratio. n = 4 adult and 1 pediatric MSC(AT). Data represent mean ± SD, comparisons done with paired Wilcoxon tests, * p < 0.05

Article Snippet: The following primary antibodies were used at the dilutions indicated: CD26 (OriGene Technologies Inc., MD, USA; diluted 1:500), β-actin (Chemicon MAB1501; diluted 1: 100,000).

Techniques: Flow Cytometry, Inhibition, Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: CD26 is a senescence marker associated with reduced immunopotency of human adipose tissue-derived multipotent mesenchymal stromal cells

doi: 10.1186/s13287-022-03026-4

Figure Lengend Snippet: Viability and CD26 surface levels in late passage MSC(AT) decrease following senolytic treatment. A Cell viability of late passage MSC(AT) that were either treated with DMSO (vehicle control) or a senolytic: NAVI: navitoclax (20 μM), QUER: quercetin (+:400 and ++:800 μM) and DMAG: 17-DMAG (+:25.6 and ++:51.2 μM). B CD26 gMFI and C %CD26 high MSCs among living MSC(AT). [ n = 4 adult and 1 pediatric LP-MSC(AT)]. Data presented as mean ± SD, comparisons done with paired one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The following primary antibodies were used at the dilutions indicated: CD26 (OriGene Technologies Inc., MD, USA; diluted 1:500), β-actin (Chemicon MAB1501; diluted 1: 100,000).

Techniques:

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: Concentration response curves of DPP-4 inhibitory activities by trelagliptin, alogliptin and sitagliptin. Activity was measured as described under Materials and Methods.

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Concentration Assay, Activity Assay

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: Progress curves at 405 nm for pNA generation were recorded over 2000 sec using a 10 sec interval. Reaction was initiated with 1 nmol/L DPP-4 in the presence of 400 μmol/L GP-pNA substrate (approximately 4x Km) and varying concentrations of trelagliptin. Inset: Replot of apparent association rate constant, k’, against trelagliptin concentration used to estimate k on ’ from the slope according to .

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: A preformed enzyme-inhibitor complex (where [DPP-4] = 50 nmol/L and trelagliptin concentration is as shown in the plot) was diluted 50-fold into a solution containing 2 mmol/L GP-pNA substrate (approximately 17x Km). Absorbance readings were taken every 10 seconds. Inset: Replot of the apparent dissociation rate constant, k’, against trelagliptin concentration used to estimate k off from the Y-intercept according to .

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: Comparison of x-ray crystal structure of inhibitors bound to DPP-4 for alogliptin (left panel) and trelagliptin (right panel).

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Comparison

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: Close-up showing potential differential interaction of F-atom of trelagliptin (right panel) with Trp659 residue in DPP-4 crystal structure as compared to H-atom of alogliptin (left panel).

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Residue

Journal: PLoS ONE

Article Title: Trelagliptin (SYR-472, Zafatek), Novel Once-Weekly Treatment for Type 2 Diabetes, Inhibits Dipeptidyl Peptidase-4 (DPP-4) via a Non-Covalent Mechanism

doi: 10.1371/journal.pone.0157509

Figure Lengend Snippet: Observed value plotted in “X” and predicted relationship between pharmacokinetics and pharmacodynamics by sigmoid Emax model is indicated by solid line. The plasma trelagliptin concentration estimated to yield 70% and 80% inhibition of human plasma DPP-4 activity was 2.31 ng/mL and 3.13 ng/mL, respectively.

Article Snippet: Recombinant human DPP-4 (Abnova, Taiwan) activity was assayed using the fluorescent substrate Gly-Pro-7-amido-4-methyl-coumarin (GP-AMC) (90 μmol/L final concentration) and carried out in pH 7.8 buffer containing 25 mmol/L HEPES, 140 mmol/L NaCl, 1 mg/mL bovine serum albumin for 15 min at 37°C.

Techniques: Concentration Assay, Inhibition, Activity Assay

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: DPP4 expression is upregulated in intestinal fibro-stenotic areas of CD patients. (A) Volcano plot showing differentially expressed genes (DEGs) between stenotic (B2) and non-stenotic (B1) CD tissues ( GSE66207 dataset). Red dots indicate DEGs with |log2FC| >1 and p < 0.05. (B, C) Hub genes identified using the MCODE and CytoHubba plugins, respectively. (D) RT-qPCR analysis of DPP4 mRNA levels in intestinal tissues. (E) Western blot analysis of DPP4 protein expression in colonic tissues from healthy controls, non-stenotic, and stenotic regions of CD patients. (F) Densitometric quantification of DPP4 protein normalised to GAPDH (corresponding to panel E). (G) Representative endoscopic images of healthy controls, non-stenotic, and stenotic intestinal regions in CD patients, illustrating macroscopic features of stricture. Corresponding immunohistochemical (IHC) images show DPP4 expression in colonic tissues from the same groups. Adjacent serial sections stained with Masson’s trichrome highlight fibrotic areas. (H) Quantification of IHC DPP4 staining (IOD/area) across groups. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Elevated DPP4 expression contributes to fibrotic remodelling in a chronic colitis model. (A) Colon length quantification in control ( n = 6) and DSS-treated ( n = 6) mice. (B) Representative H&E-stained colon sections (left) and histologic inflammation scores (right). (C) Representative Masson’s trichrome-stained colon sections and quantification of collagen volume fraction. (D) Immunofluorescence images and quantification of α -SMA⁺ fibrotic thickness in the muscularis propria. (E) qRT-PCR analysis of Col1a1 mRNA levels in colon tissues. (F) qRT-PCR analysis of Col6a1 mRNA levels in colon tissues. (G) Western blot analysis of DPP4 protein expression in control and DSS-treated mice. (H) Representative IHC staining of DPP4 expression in control and DSS-treated colons, with semiquantitative analysis. (I) Schematic overview of the chronic DSS-induced colitis model and pharmacological intervention. Mice received three cycles of 1.5% DSS (7 days per cycle), each followed by a 14-day recovery phase. The DPP4 inhibitor sitagliptin was administered via oral gavage for 21 days during the final cycle of the experiment. (J) Colon length quantification in the DSS group ( n = 6) and DSS + DPP4i group ( n = 6). (K) Representative H&E-stained colon sections (left) and histologic inflammation scores (right). (L) Representative Masson’s trichrome-stained colon sections and quantification of collagen volume fraction. (M) Immunofluorescence images and quantification of α -SMA⁺ fibrotic thickness in the muscularis propria. ( N ) RT-qPCR analysis of Col1a1 mRNA levels in colon tissues. (O) RT-qPCR analysis of Col6a1 mRNA levels in colon tissues. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Expressing, Control, Staining, Immunofluorescence, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Membrane-bound DPP4 in fibroblasts drives intestinal myofibroblast activation and migration. (A) Multiplex IHC staining of FFPE colonic sections from healthy controls, non-stenotic, and stenotic regions of CD patients. (B) Volcano plot showing DEGs in fibroblasts isolated from paired stenotic and normal CD tissues ( GSE90607 dataset, left), with DPP4 mRNA expression levels specifically highlighted (right). (C) Western blot analysis of membrane-bound DPP4 and α -SMA protein in primary HIMFs. (D) Western blot analysis and quantification of DPP4 and α -SMA expression in TGF-β–stimulated HIMFs with or without DPP4 inhibitor. (E) qRT-PCR analysis of COL1A1 mRNA expression in HIMFs. (F) qRT-PCR analysis of ACTA2 mRNA expression in HIMFs. (G) Immunofluorescence staining and quantification of Ki67⁺ proliferating HIMFs. (H) Representative images and quantification of HIMF migration in scratch wound healing assays. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Membrane, Activation Assay, Migration, Multiplex Assay, Immunohistochemistry, Isolation, Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Soluble DPP4 (sDPP4) promotes intestinal myofibroblast activation, proliferation, and migration. (A) ELISA-based quantification of soluble DPP4 (sDPP4) levels in plasma from healthy controls ( n = 10), CD patients without stenosis ( n = 20), and with stenosis ( n = 20). (B) RT-qPCR analysis of COL1A1 mRNA expression in HIMFs treated with sDPP4 in the presence or absence of DPP4 inhibitor. (C) RT-qPCR analysis of ACTA2 mRNA expression in HIMFs treated with sDPP4 in the presence or absence of DPP4 inhibitor. (D) Western blot of α -SMA protein expression in HIMFs treated with sDPP4 in the presence or absence of DPP4 inhibitor. (E) Immunofluorescence staining and quantification of Ki67⁺ proliferating HIMFs treated with sDPP4 in the presence or absence of DPP4 inhibitor. (F) Scratch wound healing assay evaluating HIMF migration at 0 and 24 hours following sDPP4 stimulation, with or without DPP4 inhibitor. Wound closure was quantified using ImageJ. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Activation Assay, Migration, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining, Wound Healing Assay

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: DPP4 drives intestinal myofibroblast activation via the PI3K-AKT pathway. (A) Western blot analysis of canonical (SMAD3) and non-canonical (PI3K-AKT, ERK, JNK, and p38) signalling pathways in TGF- β -stimulated HIMFs, treated with or without DPP4 inhibitor (sitagliptin, 20 nM). (B) Western blot analysis of p -PI3K, PI3K, p -AKT, AKT, and α -SMA expression in HIMFs treated with TGF- β , in the presence or absence of DPP4 inhibitor (20 nM) and the PI3K activator 740Y- P (10 μM). (C) Western blot analysis of p -ERK, ERK, and α -SMA expression in HIMFs treated with TGF- β , in the presence or absence of DPP4 inhibitor (20 nM) and the MEK/ERK activator C16-PAF (1 μM). (D) Scratch wound healing assay of HIMF migration at 0 and 24 hours following TGF- β stimulation with or without DPP4 inhibitor (20 nM) and the PI3K activator 740Y- P (10 μM). Wound closure was quantified using ImageJ. Ki67 immunofluorescence staining and quantification of proliferating HIMFs are shown in the lower panels. (E) Scratch wound healing assay of HIMF migration at 0 and 24 hours following TGF- β stimulation with or without DPP4 inhibitor (20 nM) and the MEK/ERK activator C16-PAF (1 μM). Wound closure was quantified using ImageJ. Ki67 immunofluorescence staining and quantification are shown in the lower panels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Activation Assay, Western Blot, Expressing, Wound Healing Assay, Migration, Immunofluorescence, Staining

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Gut microbiota-derived DPP4 is enriched in stenotic CD and associated with fibrotic remodelling. (A) Schematic overview of the faecal metagenomic sequencing strategy in controls ( n = 10), CD without stenosis ( n = 20), and CD with stenosis ( n = 20). (B) Relative abundance of microbial Dpp4 genes across the three groups, as determined by metagenomic sequencing. (C) Species-level contributions of microbial Dpp4 genes in CD patients with stenosis, showing the top five contributing species ranked by relative abundance. (D) Differential abundance of microbial Dpp4 genes assigned to four Bacteroides species in faecal metagenomes from healthy controls, non-stenotic CD, and stenotic CD patients. (E) Total faecal DPP4 enzymatic activity was measured by Gly-Pro-pNA assay in controls, CD patients without stenosis, and those with stenosis. (F) Faecal DPP4 enzymatic activity was measured in control mice and those with DSS-induced chronic colitis. (G) Time-dependent increase in DPP4 activity measured in the culture supernatants of B. thetaiotaomicron grown under anaerobic conditions. (H–I) RT-qPCR analysis of COL1A1 (H) and ACTA2 (I) mRNA levels in HIMFs treated with increasing concentrations of recombinant btDPP4 (0–400 ng/mL). (J) Western blot analysis of α -SMA protein expression in HIMFs treated with 200 ng/mL btDPP4. (K) Ki67 immunofluorescence staining and quantification of proliferating HIMFs following treatment with 200 ng/mL btDPP4. (L) Scratch wound healing assay of HIMF migration at 0 and 24 hours following treatment with 200 ng/mL btDPP4. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Derivative Assay, Sequencing, Activity Assay, Control, Quantitative RT-PCR, Recombinant, Western Blot, Expressing, Immunofluorescence, Staining, Wound Healing Assay, Migration

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Colonisation with engineered bacteria overexpressing DPP4 worsens fibrotic remodelling in DSS-induced chronic colitis. (A) Schematic illustration of engineered E. coli btDPP4 construction. (B) PCR validation of btDPP4 genomic integration in engineered E. coli strains. (C) Growth curves comparing E. coli btDPP4 and wild-type (WT) strains under standard conditions. (D) Secreted DPP4 enzymatic activity measured in the culture supernatants of E. coli btDPP4 and E. coli WT. (E) Experimental design of the chronic DSS colitis model with oral gavage of PBS, E. coli WT, or E. coli btDPP4. (F) Relative faecal E. coli load in mice treated with PBS, E. coli WT, or E. coli btDPP4. (G) Expression of btDPP4 gene in faeces from mice treated with PBS, E. coli WT, or E. coli btDPP4. (H) Faecal DPP4 enzymatic activity in mice treated with PBS, E. coli WT, or E. coli btDPP4. (I) Colon length was measured and compared across groups. (J) Representative H&E-stained colon sections and quantification of histological inflammation scores. (K) Representative Masson’s trichrome-stained sections and quantification of collagen volume fraction. (L) Immunofluorescence staining of α -SMA⁺ fibrotic areas and quantification of fibrotic thickness. (M–N) RT-qPCR analysis of Col1a1 (M) and Col6a1 ( N ) mRNA levels in mouse colon tissues. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Bacteria, Biomarker Discovery, Activity Assay, Expressing, Staining, Immunofluorescence, Quantitative RT-PCR

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Microbial-derived DPP4 inhibitor Dau-d4 suppresses intestinal myofibroblast activation. (A-B) RT-qPCR analysis of COL1A1 and ACTA2 mRNA expression in HIMFs treated with increasing concentrations of recombinant microbial DPP4 (btDPP4, hereafter referred to as mDPP4), with or without the microbial DPP4 inhibitor Dau-d4 (0–200 nM). (C) Western blot analysis of α -SMA protein expression in HIMFs stimulated with mDPP4, with or without 100 nM Dau-d4. (D) Ki67 immunofluorescence staining and quantification of proliferating HIMFs stimulated with mDPP4, with or without 100 nM Dau-d4. (E) Scratch wound healing assay showing HIMF migration at 0 and 24 hours following treatment with mDPP4, in the presence or absence of 100 nM Dau-d4. (F) Western blot analysis of p -PI3K, PI3K, p -AKT, AKT, and α -SMA expression in HIMFs treated with vehicle control, mDPP4 (200 ng/mL), mDPP4 + Dau-d4 (100 nM), or mDPP4 + Dau-d4 + PI3K activator 740Y- P (10 μM) for 24 hours. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Derivative Assay, Activation Assay, Quantitative RT-PCR, Expressing, Recombinant, Western Blot, Immunofluorescence, Staining, Wound Healing Assay, Migration, Control

Journal: Gut Microbes

Article Title: Dual-source DPP4 drives intestinal fibrosis in Crohn’s disease: synergistic therapeutic targeting of host and microbiota pathways

doi: 10.1080/19490976.2025.2593119

Figure Lengend Snippet: Dual inhibition of bacterial- and host-derived DPP4 synergistically attenuates intestinal fibrosis in vivo. (A) Schematic of the experimental design showing late-phase intervention with the microbiota-derived DPP4 inhibitor Dau-d4 (10 mg/kg), alone or in combination with the host-derived DPP4 inhibitor sitagliptin, during the final cycle of DSS-induced chronic colitis. (B) DPP4 activity was measured in the murine faeces following treatment. (C) Colon length was measured and compared across groups. (D) Representative H&E-stained colon sections and quantification of histological inflammation scores. (E) Representative images of Masson’s trichrome staining and quantification of collagen volume fraction. (F) Immunofluorescence staining of α -SMA⁺ areas and quantification. (G-H) RT-qPCR analysis of Col1a1 and Col6a1 mRNA expression in murine colonic tissues. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: After blocking, sections were incubated overnight at 4 °C with anti-DPP4 antibody (1:100, Abmart, China).

Techniques: Inhibition, Derivative Assay, In Vivo, Activity Assay, Staining, Immunofluorescence, Quantitative RT-PCR, Expressing