recombinant full length murine angptl4  (R&D Systems)

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Lipolysis products from triglyceride-rich lipoproteins induce stress protein ATF3 in osteoblasts.

doi: 10.1002/jor.25756

Figure Lengend Snippet: FIGURE 5 Lipolysis products induce the expression of Angptl4 in MC3T3‐E1 cells. Cells were exposed to 0−150 mg/dL TGRL mixed with 2 units/mL LPL for 3 or 6 h. Bars represent mean ± SEM, n = 4−6, ****p < 0.0001 compared to 0 TGRL + LPL (Ctrl). LPL, lipoprotein lipase; TGRL, triglyceride‐rich lipoproteins.

Article Snippet: Recombinant full length murine ANGPTL4 (4880‐AN‐050) was purchased from R&D Systems.

Techniques: Expressing

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Lipolysis products from triglyceride-rich lipoproteins induce stress protein ATF3 in osteoblasts.

doi: 10.1002/jor.25756

Figure Lengend Snippet: FIGURE 6 Recombinant ANGPTL4 inhibits lipolysis product‐ induction of Atf3 in MC3T3‐E1 cells. Cells were exposed to 100 mg/dL TGRL mixed with 2 units/mL LPL for 6 h. In some experiments, LPL and ANGPTL4 (0.1−5 µg/mL) were mixed for 30 min before the addition of TGRL. Ctrl represents cells not exposed to TGRL/LPL/ANGPTL4. Bars represent mean ± SEM, n = 4−6. *p < 0.05, ***p < 0.001, ****p < 0.0001 compared to 0 µg/mL ANGPTL4. LPL, lipoprotein lipase; TGRL, triglyceride‐rich lipoproteins.

Article Snippet: Recombinant full length murine ANGPTL4 (4880‐AN‐050) was purchased from R&D Systems.

Techniques: Recombinant

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Lipolysis products from triglyceride-rich lipoproteins induce stress protein ATF3 in osteoblasts.

doi: 10.1002/jor.25756

Figure Lengend Snippet: FIGURE 11 (A) Lipoprotein lipase (LPL) (or hepatic lipase) secreted by the osteoblast translocates to the plasma membrane of endothelial cells lining blood vessels, where it catalyzes the conversion of triglyceride‐rich lipoproteins (TGRL) into lipolysis products (LP) such as free fatty acids (FFA). LP translocate to the osteoblast for use as a metabolic fuel source. (B) High circulating levels of TGRL result in high levels of LP which induce an increase in ATF3 in osteoblasts (pathway 1), and induce IL6 and COX‐2, known downstream genes of ATF3; LP also induce intracellular ROS accumulation, which together result in inflammation. LP induce an increase in angiopoietin‐like 4 (ANGPTL4) (pathway 2), an inhibitor of LPL, which may act in a negative feedback loop, reducing LPL activity, reducing LP accumulation in the local environment, and protecting cells from inflammation. Note it is possible that lipases may be active on the osteoblast cell membrane itself. ATF3, activating transcription factor 3; ROS, reactive oxygen species.

Article Snippet: Recombinant full length murine ANGPTL4 (4880‐AN‐050) was purchased from R&D Systems.

Techniques: Clinical Proteomics, Membrane, Activity Assay

Journal: Poultry Science

Article Title: Single-cell RNA sequencing reveals intrahepatic signature related to pathobiology of duck hepatitis A virus type 3 (DHAV-3) infection

doi: 10.1016/j.psj.2025.104798

Figure Lengend Snippet: Dynamic response of T/NK cells during DHAV-3 infection. (A) UMAP plot of 15 subclusters of T/NK population. (B) The ratio of each T/NK subcluster in each group. (C) The boxplot representing ratio of each subcluster in R24 and S24. (D) RNA velocity analysis showing the transition potential among CD8+ T-cell subsets. (E) The correlation between viral load and PLAC8 expression level in viral RNA positive cells. (F) The qPCR results showing the PLAC8 expression and DHAV-3 viral load in multiple tissues from R24 and S24 ducklings. (G) PLAC8 overexpression promoted DHAV-3 replication in primary liver cells. The p values shown in the figures are represented by *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Recombinant plasmids pcDNA3.1- PLAC8 and pcDNA3.1- ANGPTL4 were constructed by double digestion and verified by Sanger sequencing (Sangon Biotech, China).

Techniques: Infection, Expressing, Over Expression

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Comparison and interpretation of fibroblast disease signatures from different heart failure models. A Comparing intersections of upregulated genes in different heart failure (HF) models. B Intersection quantification via Jaccard index. C Comparison of direction of regulation between studies. Pearson correlation was calculated between log fold change vectors of signature genes in pairwise comparisons. Each study comparison was based on the upregulated genes from the study on the x-axis. ** p < 0.01. D Heatmaps of gene set overrepresentation in study specific fibroblast disease signatures. Hypergeometric test with Benjamini–Hochberg correction, * q < 0.01, ** q < 0.001, *** q < 0.0001. E Estimated pathway activities with PROGENy based on effect size (log fold change) of footprint genes compared between HF models. F Expression values of selected fibrosis and inflammatory genes in individual fibroblasts in HFpEF (purple) and control (orange) mice. All genes were significantly upregulated (Wilcoxon test, adj. p value < 0.05). G Immunofluorescence images of collagen IV (red) and DAPI (blue) staining of left ventricular heart sections. Lower panels show magnifications of the areas marked by white boxes. White arrows indicate capillaries or larger blood vessels. Scale bars in the right bottom corner indicate 50 μm length. H Immunohistological staining of Angptl4 protein in left ventricular heart sections

Article Snippet: One day after plating of 300,000 cells/well in 6-well plates on hydrogels (Softwell, 50 kPa) with lower stiffness than plastic surfaces to mimic a diseased heart, cells were treated either with recombinant human ANGPTL4 (R&D, #4487-AN-050) 2 μg/ml or PBS for 24 h in fibroblast growth medium at 37 °C.

Techniques: Comparison, Expressing, Control, Immunofluorescence, Staining

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Angptl4 improves diastolic dysfunction by reducing collagen IV deposition in vivo and in vitro. A In vivo study design comparing recombinant murine Angptl4 peptide (rANGPTL4, 200 ng in 50 µl NaCl) vs. NaCl control (0.9%, 50 µl) administration every second day i.p. for 5 weeks starting after 5 weeks of dietary induction. Murine HFpEF induction by 0.5 g/L L-NAME and 60% high-fat diet for 10 weeks in total. Echocardiography (echo) captured cardiac systolic and diastolic function at baseline, after 5 (prior to i.p. injection start) and 10 weeks. Created with BioRender. B Time course of diastolic function determined by E/E’ (PW Doppler velocity across the mitral valve (E) and peak tissue Doppler at the mitral valve annulus (E’) during early diastole). n = 10/10/9/9. C Comparison of the experimental groups after 10 weeks. n = 10/10/9/9. One-way ANOVA with Tukey correction for multiple comparison or Kruskal–Wallis test according to normality determined by Shapiro–Wilk test. p values < 0.09 shown above bars, p < 0.05 defined as statistically significant. HW/TL heart weight/ tibia length, LA left atrium, LVEF left ventricular ejection fraction, LWDd left ventricular lateral wall diameter in end-diastole from short axis views D Comparison of E/E’ after 10 weeks in HFpEF mice either treated with control or rANGPTL4. n = 9/9, unpaired t test, p value shown above bar. E Experimental design (left panel) of stimulating human ventricular cardiac fibroblasts (cFBs) in vitro with 2 µg/ml human recombinant ANGPTL4 or PBS for 24 h. Resulting mRNA levels (right panel) were determined by qPCR and values indicate fold change relative to control mean. n = 12/12, unpaired t test or Mann–Whitney test according to normality determined by Shapiro–Wilk test, p values shown above bars. Created with BioRender. F Representative immunofluorescence stainings of collagen IV 1:200 (pink) and DAPI (blue). Whole heart long-axis cryo-sections depicted in the top row with white boxes indicating magnifications shown in bottom row and respective scale bars. G Quantification of F by scanning whole heart sections using a slide scanner and semi-automated analysis of the whole left ventricular (LV) tissue by normalizing the collagen IV positive LV area to total LV area using QuPath. One-way ANOVA, p values < 0.05 shown above bars

Article Snippet: One day after plating of 300,000 cells/well in 6-well plates on hydrogels (Softwell, 50 kPa) with lower stiffness than plastic surfaces to mimic a diseased heart, cells were treated either with recombinant human ANGPTL4 (R&D, #4487-AN-050) 2 μg/ml or PBS for 24 h in fibroblast growth medium at 37 °C.

Techniques: In Vivo, In Vitro, Recombinant, Control, Injection, Comparison, MANN-WHITNEY, Immunofluorescence

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Plasma ANGPTL4 is increased in HFpEF patients. A Circulating levels of ANGPTL4 in human plasma samples of HFpEF and age-matched controls measured by sandwich ELISA. n = 19/20, Mann–Whitney U test, * p < 0.05. B) ANGPTL4 plasma levels in relation to NYHA functional class of all recruited patients. ANOVA, p value < 0.05, n = 10/21/3 in baseline and n = 11/18/5 in 12 months (12 M) follow-up. C Correlation of clinical parameters to ANGPTL4 circulating levels in all patients (control and HFpEF) and D as subanalysis only in HFpEF patients using simple linear regression. p-val indicates uncorrected p value. hs high sensitivity, LA left atrial, MFU months follow-up, SVES supraventricular extrasystoles. Plots in ( A , B ) display mean ± SD

Article Snippet: One day after plating of 300,000 cells/well in 6-well plates on hydrogels (Softwell, 50 kPa) with lower stiffness than plastic surfaces to mimic a diseased heart, cells were treated either with recombinant human ANGPTL4 (R&D, #4487-AN-050) 2 μg/ml or PBS for 24 h in fibroblast growth medium at 37 °C.

Techniques: Clinical Proteomics, Sandwich ELISA, MANN-WHITNEY, Functional Assay, Control

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Comparison and interpretation of fibroblast disease signatures from different heart failure models. A Comparing intersections of upregulated genes in different heart failure (HF) models. B Intersection quantification via Jaccard index. C Comparison of direction of regulation between studies. Pearson correlation was calculated between log fold change vectors of signature genes in pairwise comparisons. Each study comparison was based on the upregulated genes from the study on the x-axis. ** p < 0.01. D Heatmaps of gene set overrepresentation in study specific fibroblast disease signatures. Hypergeometric test with Benjamini–Hochberg correction, * q < 0.01, ** q < 0.001, *** q < 0.0001. E Estimated pathway activities with PROGENy based on effect size (log fold change) of footprint genes compared between HF models. F Expression values of selected fibrosis and inflammatory genes in individual fibroblasts in HFpEF (purple) and control (orange) mice. All genes were significantly upregulated (Wilcoxon test, adj. p value < 0.05). G Immunofluorescence images of collagen IV (red) and DAPI (blue) staining of left ventricular heart sections. Lower panels show magnifications of the areas marked by white boxes. White arrows indicate capillaries or larger blood vessels. Scale bars in the right bottom corner indicate 50 μm length. H Immunohistological staining of Angptl4 protein in left ventricular heart sections

Article Snippet: For murine recombinant ANGPTL4 (R&D, 4880-AN-050) peptide treatment, HFpEF was induced as described above and after 5 weeks i.p. injections of either 200 ng rANGPTL4 (in 50 μl NaCl) or NaCl (50 μl) as control.

Techniques: Comparison, Expressing, Control, Immunofluorescence, Staining

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Angptl4 improves diastolic dysfunction by reducing collagen IV deposition in vivo and in vitro. A In vivo study design comparing recombinant murine Angptl4 peptide (rANGPTL4, 200 ng in 50 µl NaCl) vs. NaCl control (0.9%, 50 µl) administration every second day i.p. for 5 weeks starting after 5 weeks of dietary induction. Murine HFpEF induction by 0.5 g/L L-NAME and 60% high-fat diet for 10 weeks in total. Echocardiography (echo) captured cardiac systolic and diastolic function at baseline, after 5 (prior to i.p. injection start) and 10 weeks. Created with BioRender. B Time course of diastolic function determined by E/E’ (PW Doppler velocity across the mitral valve (E) and peak tissue Doppler at the mitral valve annulus (E’) during early diastole). n = 10/10/9/9. C Comparison of the experimental groups after 10 weeks. n = 10/10/9/9. One-way ANOVA with Tukey correction for multiple comparison or Kruskal–Wallis test according to normality determined by Shapiro–Wilk test. p values < 0.09 shown above bars, p < 0.05 defined as statistically significant. HW/TL heart weight/ tibia length, LA left atrium, LVEF left ventricular ejection fraction, LWDd left ventricular lateral wall diameter in end-diastole from short axis views D Comparison of E/E’ after 10 weeks in HFpEF mice either treated with control or rANGPTL4. n = 9/9, unpaired t test, p value shown above bar. E Experimental design (left panel) of stimulating human ventricular cardiac fibroblasts (cFBs) in vitro with 2 µg/ml human recombinant ANGPTL4 or PBS for 24 h. Resulting mRNA levels (right panel) were determined by qPCR and values indicate fold change relative to control mean. n = 12/12, unpaired t test or Mann–Whitney test according to normality determined by Shapiro–Wilk test, p values shown above bars. Created with BioRender. F Representative immunofluorescence stainings of collagen IV 1:200 (pink) and DAPI (blue). Whole heart long-axis cryo-sections depicted in the top row with white boxes indicating magnifications shown in bottom row and respective scale bars. G Quantification of F by scanning whole heart sections using a slide scanner and semi-automated analysis of the whole left ventricular (LV) tissue by normalizing the collagen IV positive LV area to total LV area using QuPath. One-way ANOVA, p values < 0.05 shown above bars

Article Snippet: For murine recombinant ANGPTL4 (R&D, 4880-AN-050) peptide treatment, HFpEF was induced as described above and after 5 weeks i.p. injections of either 200 ng rANGPTL4 (in 50 μl NaCl) or NaCl (50 μl) as control.

Techniques: In Vivo, In Vitro, Recombinant, Control, Injection, Comparison, MANN-WHITNEY, Immunofluorescence

Journal: Basic Research in Cardiology

Article Title: Single-cell transcriptomics reveal distinctive patterns of fibroblast activation in heart failure with preserved ejection fraction

doi: 10.1007/s00395-024-01074-w

Figure Lengend Snippet: Plasma ANGPTL4 is increased in HFpEF patients. A Circulating levels of ANGPTL4 in human plasma samples of HFpEF and age-matched controls measured by sandwich ELISA. n = 19/20, Mann–Whitney U test, * p < 0.05. B) ANGPTL4 plasma levels in relation to NYHA functional class of all recruited patients. ANOVA, p value < 0.05, n = 10/21/3 in baseline and n = 11/18/5 in 12 months (12 M) follow-up. C Correlation of clinical parameters to ANGPTL4 circulating levels in all patients (control and HFpEF) and D as subanalysis only in HFpEF patients using simple linear regression. p-val indicates uncorrected p value. hs high sensitivity, LA left atrial, MFU months follow-up, SVES supraventricular extrasystoles. Plots in ( A , B ) display mean ± SD

Article Snippet: For murine recombinant ANGPTL4 (R&D, 4880-AN-050) peptide treatment, HFpEF was induced as described above and after 5 weeks i.p. injections of either 200 ng rANGPTL4 (in 50 μl NaCl) or NaCl (50 μl) as control.

Techniques: Clinical Proteomics, Sandwich ELISA, MANN-WHITNEY, Functional Assay, Control

Journal: Agriculture Communications

Article Title: Single-cell transcriptional profiling of porcine muscle satellite cells and myoblasts during myogenesis

doi: 10.1016/j.agrcom.2024.100026

Figure Lengend Snippet: Fig. 7. Integrated analysis of publicly-available murine single cell RNA-sequencing (scRNA-seq) data and in vitro treatment with recombinant ANGPTL4 protein. (a) Integrated analysis of murine skeletal muscle scRNA-seq data, clustering of cell types, and analysis of cell–cell communication networks. (b) and (d), EdU staining of porcine MuSCs and C2C12 cells. The S-phase is stained with EdU (green), and the nuclei are stained with Hoechst 33342 (blue). (c) and (e), Immunofluorescence staining of MyHc. Porcine myofibers are stained green, and nuclei are stained blue. Abbreviations: ANGPTL4, angiopoietin like4; EdU, 5-ethynyl-20-deoxyuridine; MyHc, myosin heavy chain; FAPs, fibro-adipogenic progenitors.

Article Snippet: Briefly, MuSCs were seeded into 96-well plates and treated with DMEM/F-12 proliferation medium containing 2.5 μg/mL recombinant ANGPTL4 protein (CUSABIO, Wuhan, China) for two days [17].

Techniques: RNA Sequencing, In Vitro, Recombinant, Staining