Journal: bioRxiv

Article Title: Simultaneous and sequential multi-species coronavirus vaccination

doi: 10.1101/2022.05.07.491038

Figure Lengend Snippet: (A) Schematics of mRNA vaccine construct design against pathogenic human coronavirus species. Each construct has regulatory elements (5’UTR, 3’UTR and polyA) and spike ORF. The domain structures as well as engineered mutations of translated spike proteins of SARS-CoV-2 Delta variant (Delta), SARS-CoV (SARS) and MERS-CoV (MERS). (B) Engineered mutations in spike protein structures of SARS-CoV-2 Delta, SARS-CoV and MERS-CoV. The N-terminal domain (NTD, blue), receptor binding domain (RBD, green) and S2 subunit (orange) of one protomer along with homologous HexaPro mutations (pink) and Delta variant mutations (red) were highlighted in the spike trimer structures. (C) Schematics of characterization of LNP-mRNA vaccine formulations. Assembly procedure of LNP-mRNA vaccine on NanoAssemblr Ignite and downstream biophysical characterization assays. (D) Histogram displaying radius distribution of LNP-mRNA formulations of SARS-CoV-2 Delta and a Triplex (Delta + SARS + MERS) (abbreviated as Triplex-CoV or Triplex), measured by dynamic light scattering (DLS). The polydispersity index and mean radius of each LNP sample were shown at top left corner. (E) Transmission electron microscope (TEM) images of Delta and Triplex-CoV LNP-mRNAs. (F) Surface expression of functional spike proteins in 293T cells after electroporation of corresponding mRNA, as detected by human ACE2 or human DPP4 Fc fusion protein bound to PE anti-Fc antibody. (G) Schematics of vaccination schedule of the Triplex LNP-mRNA formulations, as well as downstream assays to evaluate the antibody responses and other immunological profiles. (H) Binding antibody titers of plasma samples from mice administered with PBS or different LNP-mRNAs (n = 9) against RBD or ectodomain (ECD) of SARS-CoV-2 wild type (WT, Wuhan/WA-1), Delta variant, SARS and MERS spikes. The binding antibody titers were quantified by area under curve of log 10 -transformed titration curve (log 10 AUC) in . The mice were intramuscularly injected with two doses (x2, 2 weeks apart) of PBS, 1μg SARS-CoV-2 Delta variant LNP-mRNA (delta), 1μg or 3μg equal mass mixture of Delta, SARS and MERS LNP-mRNA (Triplex-CoV). Notes: In the dot-box plots of this figure, each dot represents data from one mouse. Data are shown as mean ± s.e.m. plus individual data points in plots. Two-way ANOVA with Tukey’s multiple comparisons test was used to assess statistical significance. Statistical significance labels: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Non-significant comparisons are not shown, unless otherwise noted as n.s., not significant.

Article Snippet: To detect surface-protein expression, the cells were stained with ACE2–Fc chimera (Genscript, Z03484) or DPP4-Fc (Sino Biological, 10688-H01H) in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA) for 30 min on ice.

Techniques: Construct, Variant Assay, Binding Assay, Transmission Assay, Microscopy, Expressing, Functional Assay, Electroporation, Transformation Assay, Titration, Injection

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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: DPP4 activity in human and murine faecal samples was quantified using Gly-Pro- p -nitroanilide hydrochloride (Gly-Pro-pNA, TargetMol, USA) as the substrate.

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

Journal: Viruses

Article Title: Phage-Displayed Mimotopes of SARS-CoV-2 Spike Protein Targeted to Authentic and Alternative Cellular Receptors

doi: 10.3390/v14020384

Figure Lengend Snippet: Characteristics of recombinant human cell receptors and viral spike proteins.

Article Snippet: hDPP4 , Sino Biological , 10688-H08H , D34 , P766 , 744 , 86.3.

Techniques: Recombinant, Molecular Weight

Journal: Pathogens and Disease

Article Title: Protective vascular coagulation in response to bacterial infection of the kidney is regulated by bacterial lipid A and host CD147

doi: 10.1093/femspd/fty087

Figure Lengend Snippet: Epithelial signaling factors induce endothelial TF activation. ( A ) Proteome array of supernatant of epithelial cells infected with CFT073 for 4 h. Supernatant from uninfected cells was used as the comparative control. Bars represent index of mean pixel density from the nitrocellulose membranes, n = 3. ( B ) CD26 and ( C ) CD147 expression in the supernatant of RPTEC/TERT1 cells stimulated for 4 h with cell culture medium, 5 mg/ml LPS or infected at an MOI 1:20 with CFT073 or LT002. Measurements were performed by ELISA, n = 5. ( D ) TF pro-coagulant activity, pM, measured using chromogenic assays in endothelial cells treated with the noted concentrations of ( D ) CD26, or ( E ) CD147, n = 3. All bars represent mean ± SD.

Article Snippet: Endothelial cells were stimulated with homo-dimer human recombinant EMMPRIN (Abcam, Sweden #AB155636) or human CD26/DPP4 recombinant protein (Sino Biological #10 688-HNCH-10) for 4 h before TF activation determination at the noted concentrations.

Techniques: Activation Assay, Infection, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin improves left ventricular relaxation in obese ZSF1 rats. Plasma DPP‐4 activity (A), aGIP levels (B), fasting glucose levels (C), bodyweight (D), mitral valve deceleration time (E) and E/E’ ratio (F) in 20‐wk‐old linagliptin‐ (Obese + Lina) and placebo‐treated obese (Obese) ZSF1 rats (n = 7 per group). aGIP, active glucose‐dependent insulinotropic peptide; DPP‐4, dipeptidyl peptidase‐4; E, early mitral inflow peak velocity; E’, early diastolic mitral annulus peak velocity; ND, not detected; RFU, relative fluorescence units. Data are expressed as mean ± SEM. Data were analysed using a two‐tailed unpaired Student t test, except D was analysed by a Mann‐Whitney U test. * Indicates P < .05, ** P < .01 and *** P < .001

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: Activity Assay, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin decreases F passsive by increasing titin phosphorylation at N2Bus S4010 in human cardiomyocytes in vitro. A, Cardiomyocyte passive stiffness in human cardiomyocytes treated with 300 ng/mL DPP‐4, 100 nmol/L linagliptin and 300 ng/mL DPP‐4, or control (DMSO/PBS) for 30 min (n = 3 different left ventricular tissues measuring at least 12 cardiomyocytes from each left ventricular tissue per condition). B, PKA‐mediated N2Bus S4010 phosphorylation and representative Coomassie Blue stained‐PVDF membranes and (C) PKG‐dependent N2Bus S4099 phosphorylation in human cardiomyocytes (n = 5 different left ventricular tissues) treated with vehicle control (DMSO/PBS), 300 ng/mL DPP‐4, 300 ng/mL DPP‐4 and 100 nmol/L linagliptin, or 100 nmol/L linagliptin for 2 h. F passive , passive stiffness; N2Bus, N2B unique sequence. Panel A was analysed using a two‐tailed unpaired Student t test with * P < .05. Panel B and C were analysed using a one‐way ANOVA with Dunnett's multiple comparison post hoc test with * P < .05 and *** P < .001 comparing control, DPP‐4 + Lina and Lina to DPP‐4

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: In Vitro, Staining, Sequencing, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin prevents DPP‐4‐mediated titin cleavage in human cardiomyocytes in vitro. A‐B, Western blots of total titin, containing the two isoforms N2BA and N2B and the known degradation product titin‐2 (T2) of human intact non‐permeabilized (A) and non‐intact permeabilized (B) human cardiomyocytes exposed to control PBS/DMSO (Ctrl) for 0 or 2 h (lane 1 and 2, respectively), 300 ng/mL DPP‐4 for 30 min and 2 h (lane 3 and 4, respectively), 300 ng/mL DPP‐4 and 100 nmol/L linagliptin for 2 h (lane 5) or 100 nmol/L linagliptin alone for 2 h (lane 6) in vitro (n = 3 different left ventricular tissues). Titin cleavage is indicated by red asterisks. Western blots of the specific N2Bus region (C; cross‐species conserved sequence QELLSKETLFP) and PEVK region (D; cross‐species conserved sequence KLRPGSGGEKPP) of human cardiomyocytes exposed to control PBS/DMSO (Ctrl) for 0 or 2 h (lane 1 and 2, respectively), 300 ng/mL DPP‐4 for 30 min and 2 h (lane 3 and 4, respectively), 300 ng/mL DPP‐4 and 100 nmol/L linagliptin for 2 h (lane 5) or 100 nmol/L linagliptin alone for 2 h (lane 6) (n = 3 different left ventricular tissues). Titin cleavage is indicated by red asterisks. The framed area represents the location for the zoomed images presented on the right side. Ctrl, control; DPP‐4, dipeptidyl peptidase‐4; F passive , passive stiffness; Lina, linagliptin; N2Bus, N2B unique sequence. Data are expressed as mean ± SEM

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: In Vitro, Western Blot, Sequencing