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

Journal: BMJ Open Diabetes Research & Care

Article Title: Inflammation increases plasma angiopoietin-like protein 4 in patients with the metabolic syndrome and type 2 diabetes

doi: 10.1136/bmjdrc-2014-000034

Figure Lengend Snippet: Clinical characteristics

Article Snippet: Briefly, 96-well plates were coated with antihuman ANGPTL4 polyclonal goat IgG antibody (AF3485, R&D Systems) and incubated overnight at 4°C.

Techniques:

Journal: BMJ Open Diabetes Research & Care

Article Title: Inflammation increases plasma angiopoietin-like protein 4 in patients with the metabolic syndrome and type 2 diabetes

doi: 10.1136/bmjdrc-2014-000034

Figure Lengend Snippet: Plasma angiopoietin-like protein 4 (ANGPTL4) in healthy controls (Controls), patients with metabolic syndrome without (MetS−I) and with low-grade inflammation (MetS+I) and patients with type 2 diabetes mellitus (T2DM). Plasma was taken from Controls, patients with MetS−I, MetS+I and T2DM, and plasma ANGPTL4 levels were determined. As compared with plasma ANGPTL4 levels (median (IQR)) in Controls (3.7 (2.8–5.2) ng/mL), ANGPTL4 levels are increased in patients with MetS−I (4.6 (3.3–6.5) ng/mL, p=0.006), patients with MetS+I (6.0 (3.8–7.7) ng/mL, p<0.001) and patients with T2DM (6.4 (5.2–8.2) ng/mL, p<0.001). Furthermore, ANGPTL4 was increased in T2DM compared with MetS−I (p<0.001). Significant as compared with Controls. *p<0.001, #p<0.05.

Article Snippet: Briefly, 96-well plates were coated with antihuman ANGPTL4 polyclonal goat IgG antibody (AF3485, R&D Systems) and incubated overnight at 4°C.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Inflammation increases plasma angiopoietin-like protein 4 in patients with the metabolic syndrome and type 2 diabetes

doi: 10.1136/bmjdrc-2014-000034

Figure Lengend Snippet: Correlation between plasma C reactive protein (CRP) and angiopoietin-like protein 4 (ANGPTL4). Plasma was taken from healthy controls, patients with metabolic syndrome without and with low-grade inflammation and patients with type 2 diabetes mellitus, and plasma ANGPTL4 levels as well as CRP levels were determined. Plasma CRP correlates positively with plasma ANGPTL4 (r=0.295, p<0.001).

Article Snippet: Briefly, 96-well plates were coated with antihuman ANGPTL4 polyclonal goat IgG antibody (AF3485, R&D Systems) and incubated overnight at 4°C.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Inflammation increases plasma angiopoietin-like protein 4 in patients with the metabolic syndrome and type 2 diabetes

doi: 10.1136/bmjdrc-2014-000034

Figure Lengend Snippet: Inflammatory stimuli increase angiopoietin-like protein 4 (ANGPTL4) gene and protein expression in human macrophages. (A) Human THP-1 macrophages were incubated with various Toll-like receptor (TLR) agonists for 12 h, and changes in expression of ANGPTL4, Gdf15 and Cxcl2 were determined. (B) Human THP-1 macrophages were incubated with various TLR agonists for 24 h, and ANGPTL4 levels in medium were assessed. (C) Human U937 macrophages were incubated with lipopolysaccharide (LPS) (100 ng/mL; 4 h) and ANGPTL4 mRNA expression was determined; human monocyte-derived macrophages were incubated with LPS (1 µg/mL; 24 h) and ANGPTL4 mRNA expression was determined. (D and E) THP-1 macrophages were incubated with LPS, intralipid, or both for 24 h and ANGPTL4 mRNA expression (D) or protein secretion (E) were determined. Values are means±SEM (n=3–6). Differences as compared with controls were evaluated by Student t test. *p<0.05.

Article Snippet: Briefly, 96-well plates were coated with antihuman ANGPTL4 polyclonal goat IgG antibody (AF3485, R&D Systems) and incubated overnight at 4°C.

Techniques: Expressing, Incubation, Derivative Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Exploring Memory Function Beyond Immune Cells: ANGPTL4-Mediated Memory Functions in Tissue Resident Stem Cells.

doi: 10.1002/advs.202307545

Figure Lengend Snippet: Figure 6. Epigenetic modifications contribute to memory function regulation following foreign antigen exposure through ANGPTL4. We hypothesized that adverse effect of foreign antigen exposures on endometrial stem cells could be mediated through site-specific histone H3 methylation. We then fur- ther determined whether ANGPTL4 as an upstream regulator could modulate these epigenetic modifications A). Endometrial stem cells were treated with 𝛽-glucan (25 μg ml−1) alone or were simultaneously transfected with shRNA targeting ANGPTL4. Subsequently, attenuating effects of ANGPTL4 knock- down on modifications of mono-, di-, and tri-methylation patterns of H3K4 (H3K4me1, H3K4me2, and H3K4me3), H3K9 (H3K9me2 and H3K9me3), and H3K27 (H3K27me1) were assessed using histone H3 modification multiplex assay kit B). Endometrial stem cells were pretreated with BIX-01294, a selective inhibitor of G9a histone methyltransferase (5 μM) for 24 h prior to additional treatment with 𝛽-glucan (25 μg ml−1) and subsequent changes in their self-renewal capability were assessed using MTT assays C). After pre-treating endometrial stem cells with BIX-01294 as previously described and subjecting them to subsequent 𝛽-glucan exposure under identical conditions mentioned earlier, we measured subsequent alterations in migratory capacity using the transwell assay D) and conducted western blotting to assess levels of MMP-2 and MMP-9 E), respectively. Abolishing effects of BIX- 01294 pretreatment on the multilineage differentiation potential of endometrial stem cells into adipocytes F) and osteoblasts G) were assessed through oil red O staining and alizarin red S staining, respectively. Effects of BIX-01294 pretreatment following 𝛽-glucan exposure on mRNA levels of several pluripotency/stemness-related genes (C-MYC, KLF4, NANOG, OCT4, and SOX2) were analyzed using real-time PCR H). 𝛽-actin was used as an internal control. PPIA was used as a housekeeping gene for real-time PCR analysis. All experiments were performed in triplicates. Data are presented as mean ± standard deviation (SD). *, p < 0.05; **, p < 0.005; and ***, p < 0.001 (two-sample t-test).

Article Snippet: Both BALB/c (were purchased from Daehan Bio Link) and ANGPTL4 KO (were purchased from Cyagen Biosciences Inc) mice were randomly divided four groups.

Techniques: Methylation, Transfection, shRNA, Knockdown, Multiplex Assay, Transwell Assay, Western Blot, Staining, Real-time Polymerase Chain Reaction, Control, Standard Deviation

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Exploring Memory Function Beyond Immune Cells: ANGPTL4-Mediated Memory Functions in Tissue Resident Stem Cells.

doi: 10.1002/advs.202307545

Figure Lengend Snippet: Figure 10. ANGPTL4 knockout results in significant alleviation of endometrial stem cell memory functions in response consecutive antigen exposure in vivo. A schematic diagram illustrating the overall experimental protocol as described in the section of “Experimental Section” is presented A). ANGPTL4 knockout (K.O) mice received intravenous treatment with 𝛽-glucan (20 mg kg−1) twice with a 7-day resting period between treatments. Endometrial stem cells were then isolated from endometrial tissues using our collagenase-based primary culture method. After isolation, mouse endometrial stem cells were cultured in vitro either under consecutive 𝛽-glucan (25 μg ml−1) exposure or non-𝛽-glucan exposure condition to properly mimic the in vivo environment of successive antigen exposures. Subsequently, attenuating effects of ANGPTL4 knockout on the self-renewal capacity of endometrial stem cells were assessed using MTT assays following successive 𝛽-glucan exposures in vivo B). Effects of consecutive 𝛽-glucan exposures on the percentage of Ki-67-positive cells were quantified using flow cytometry C). The mitigating impact of ANGPTL4 knockout on the migratory capacity of endometrial stem cells in response to 𝛽-glucan exposures in vivo was assessed using a transwell assay D). Protein levels of positive regulatory factors of cell migration (MMP-2/9) in response to 𝛽-glucan exposure with or without ANGPTL4 knockout in vivo were analyzed by western blotting E). Following consecutive 𝛽- glucan exposure, attenuating effects of ANGPTL4 knockout on the multilineage differentiation potential of endometrial stem cells into adipocytes F) and osteoblasts G) in vivo were assessed through oil red O staining and alizarin red S staining, respectively. Mitigating effects of ANGPTL4 knockout following 𝛽-glucan exposure on mRNA levels of several pluripotency/stemness-related genes (C-MYC, KLF4, NANOG, OCT4, and SOX2) in vivo were analyzed using real-time PCR H). 𝛽-actin was used as an internal control. PPIA was used as a housekeeping gene for real-time PCR analysis. All experiments were performed in triplicates. Data are presented as mean ± standard deviation (SD). *, p < 0.05; **, p < 0.005; and ***, p < 0.001 (two-sample t-test).

Article Snippet: Both BALB/c (were purchased from Daehan Bio Link) and ANGPTL4 KO (were purchased from Cyagen Biosciences Inc) mice were randomly divided four groups.

Techniques: Knock-Out, In Vivo, Isolation, Cell Culture, In Vitro, Cytometry, Transwell Assay, Migration, Western Blot, Staining, Real-time Polymerase Chain Reaction, Control, Standard Deviation

Journal: bioRxiv

Article Title: Brain endothelial PRMT5-ANGPTL4 axis regulates cerebellar inhibitory synaptogenesis and motor coordination

doi: 10.1101/2025.11.18.688985

Figure Lengend Snippet: (A) UMAP visualization of scRNA-seq data from cerebellar ECs in P20 Prmt5 fl/+ and Prmt5 fl/fl mice, with colors corresponding to the identified clusters. (B) Dot plots showing the expression of specific marker genes in each cluster. (C) Count of significant DEGs in each cluster, with upregulation indicated in orange and downregulation in blue (average |log2FC| > 0.5, adjusted p-value < 0.05). (D) Representative gene ontology (cellular component) analysis of endothelial clusters. Dot size represents the overlap proportion between DEGs and reference gene sets; color intensity reflects -log10 (q-value). (E) Volcano plot showing DEGs encoding ECM components across 6 endothelial clusters. The top 5 genes with the greatest expression differences are labeled. (F) Venn diagram showing intersections among the top 5 genes across 6 endothelial clusters, with overlaps representing shared gene counts. (G) qPCR analysis of Angptl4 mRNA levels in brain ECs derived from P20 Prmt5 fl/+ and Prmt5 fl/fl mice. **P < 0.01 (mean ± SEM, n = 3 per group). (H) Western blot analysis of ANGPTL4 protein expression in brain ECs derived from P20 Prmt5 fl/+ and Prmt5 fl/fl mice. (I) ELISA analysis of secreted ANGPTL4 protein levels. **P < 0.01 (mean ± SEM, n = 3 per group). (J) ChIP-qPCR analysis of PRMT5 occupancy on the Angptl4 promoter region in bEnd.3 cells. ****P < 0.0001 (mean ± SEM, n = 3 per group). (K) qPCR analysis of Prmt5 and Angptl4 mRNA levels in bEnd.3 cells transfected with siNC or siPrmt5 . ***P < 0.001 (mean ± SEM, n = 3 per group). (L) Western blot analysis of PRMT5 and ANGPTL4 protein levels in bEnd.3 cells transfected with siNC or siPrmt5 . GAPDH served as the loading control. (M-O) ChIP-qPCR analysis of the enrichment of H3R8me2s (M), H4R3me2s (N) and H3K9ac (O) at the Angptl4 promoter region in bEnd.3 cells transfected with siNC or siPrmt5 . **P < 0.01, ****P < 0.0001 (mean ± SEM, n = 3 per group).

Article Snippet: To measure extracellular ANGPTL4 protein levels, culture medium collected from day 4 to day 6 was analyzed using a Mouse Angiopoietin-like 4 (ANGPTL4) ELISA Kit (CUSABIO, CSB-EL001712MO) according to the manufacturer’s instructions.

Techniques: Expressing, Marker, Labeling, Derivative Assay, Western Blot, Enzyme-linked Immunosorbent Assay, ChIP-qPCR, Transfection, Control

Journal: bioRxiv

Article Title: Brain endothelial PRMT5-ANGPTL4 axis regulates cerebellar inhibitory synaptogenesis and motor coordination

doi: 10.1101/2025.11.18.688985

Figure Lengend Snippet: (A) Confocal images of VGAT (green) and Calbindin (purple) immunostaining in the cerebellar cortex of P60 Prmt5 fl/+ ;Angptl4 fl/+ , Prmt5 fl/fl ;Angptl4 fl/+ , Prmt5 fl/+ ;Angptl4 fl/fl , and Prmt5 fl/fl ;Angptl4 fl/fl mice. Scale bar, 20 µm. (B and C) Quantification of VGAT puncta number in the molecular layer (B) and surrounding individual PC bodies (C). *P < 0.05, **P < 0.01, ****P < 0.0001 (mean ± SEM, n = 5 mice per group). (D) Representative sIPSC traces recorded from PCs of P60 Prmt5 fl/+ ;Angptl4 fl/+ , Prmt5 fl/fl ;Angptl4 fl/+ , and Prmt5 fl/fl ;Angptl4 fl/fl mice. (E and F) Cumulative probability distributions and quantifications of sIPSC frequency (E) and amplitude (F). **P < 0.01, n.s., not significant (mean ± SEM, n = 6 PCs in Prmt5 fl/+ ;Angptl4 fl/+ group, n = 9 PCs in Prmt5 fl/fl ;Angptl4 fl/+ group, n = 7 PCs in Prmt5 fl/fl ;Angptl4 fl/fl group, 4 mice per group).

Article Snippet: To measure extracellular ANGPTL4 protein levels, culture medium collected from day 4 to day 6 was analyzed using a Mouse Angiopoietin-like 4 (ANGPTL4) ELISA Kit (CUSABIO, CSB-EL001712MO) according to the manufacturer’s instructions.

Techniques: Immunostaining

Journal: bioRxiv

Article Title: Brain endothelial PRMT5-ANGPTL4 axis regulates cerebellar inhibitory synaptogenesis and motor coordination

doi: 10.1101/2025.11.18.688985

Figure Lengend Snippet: (A) Representative footprints and quantification of stride length in P60 Prmt5 fl/+ ;Angptl4 fl/+ , Prmt5 fl/fl ;Angptl4 fl/+ , and Prmt5 fl/fl ;Angptl4 fl/fl mice. Red, forepaws; green, hindpaws. ****P < 0.0001 (mean ± SEM, n = 10 mice per group). (B) Quantification of latency time spent on the rotarod. Data are from four independent trials. *P < 0.05, ****P < 0.0001 (mean ± SEM, n = 10 mice per group). (C) Quantification of time spent crossing the balance beam. Data are from three independent trials. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (mean ± SEM, n = 10 mice per group).

Article Snippet: To measure extracellular ANGPTL4 protein levels, culture medium collected from day 4 to day 6 was analyzed using a Mouse Angiopoietin-like 4 (ANGPTL4) ELISA Kit (CUSABIO, CSB-EL001712MO) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: (A) Venn diagram showing 123 proteins express in both, 41 and 112 proteins uniquely express in Group A patients (CCA with ≤ 1 year survival time) and Group B patients (CCA with >1 year survival time). (B) Flow diagram of selection of candidate secreted proteins in Group A and Group B patients. Proteins identify in serum/plasma database and having signal peptide or non-classical proteins as candidate secreted proteins. ANGPTL4 is selected from Group B patients which express highest signal intensity of LC-MS/MS among candidate secreted proteins.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Selection, Clinical Proteomics, Liquid Chromatography with Mass Spectroscopy

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: (A) Representative immunohistochemical staining of ANGPTL4 in CCA tissues (magnification, x400). Hepatocytes (H) strongly express ANGPTL4 while normal bile duct (NBD) express weak positive staining and CCA cells express strongly positive staining. (B) H-scores of ANGPTL4 in normal and CCA tissues using immunohistochemistry. Mann-Whitney U -test was used to compare H-scores between normal and cancerous tissues of ANGPTL4. Statistical significance ( p < 0.05).

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, MANN-WHITNEY

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: Association between ANGPTL4 expression and clinicopathological data of CCA patients.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Expressing

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: (A) Serum ANGPTL4 level of HC and CCA patients by dot blot assay. The median ± quartile deviation, 0.1634 ± 0.1739 AU (0.0193–0.9424 AU), in healthy control and 0.6017 ± 0.3767 AU (0.0984–2.9526 AU) in CCA patients' group. To normalize the expression level, the ratio of the intensity of each dot blot to that of the positive control (pooled CCA serum) was calculated and determined as the arbitrary unit (AU). The difference of serum ANGPTL4 level between HC and CCA by Mann-Whitney U-test. (B) ROC curve of serum ANGPTL4 level as a potential biomarker for prediction of CCA in comparison group of HC, sensitivity = 80.0% and specificity = 72.7% (AUC = 0.825, 95% CI 0.751–0.899, p < 0.0001). Statistical significance ( p < 0.05).

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Dot Blot, Control, Expressing, Positive Control, MANN-WHITNEY, Biomarker Discovery, Comparison

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: Association between serum ANGPTL4 level and clinicopathological parameters in CCA patients.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques:

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: ROC curve analysis of biomarkers (ANGPTL4, CA 19-9, and CEA) for prediction of CCA in comparison group of (A) No vascular invasion vs. Vascular invasion, (B) No lymph node metastasis vs. Lymph node metastasis. Bold values * Statistical significance ( p < 0.05).

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Comparison

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: Predictive values of serum ANGPTL4, CA 19-9, and CEA levels for vascular invasion and lymph node metastasis of CCA patients, based on the optimal cutoff derived from ROC analysis and YI calculation.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques: Derivative Assay

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: Univariate and multivariate regression analysis of clinical parameters associated with vascular invasion.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques:

Journal: Frontiers in Public Health

Article Title: Serum Angiopoietin-Like Protein 4: A Potential Prognostic Biomarker for Prediction of Vascular Invasion and Lymph Node Metastasis in Cholangiocarcinoma Patients

doi: 10.3389/fpubh.2022.836985

Figure Lengend Snippet: Univariate and multivariate regression analysis of clinical parameters associated with lymph node metastasis.

Article Snippet: To investigate the specificity of the ANGPTL4 antibody, 15 μg protein each randomly selected five of healthy controls and of CCA patients was separated on a 12.5% SDS-PAGE gel at 150 V for 2 h and electrically transferred to a polyvinylidenedifluoride (PVDF) membrane at 90 V for 2 h. The membrane was blocked for 1 h using 5% skim milk in Tris-buffered saline with 0.1% Tween-20 (1X TBST, pH 7.4) before being incubated overnight at 4°C with 1:2,000 ANGPTL4 (orb353642, Biorbyt, UK).

Techniques:

Journal: Cell reports

Article Title: ANGPTL3 orchestrates hepatic fructose sensing and metabolism

doi: 10.1016/j.celrep.2025.115962

Figure Lengend Snippet: (A) Diagram of the experimental design. Male mice were fed with chow or MASLD diet for 6 weeks. Primary hepatocytes were isolated and cultured for 48 h. Conditioned media were collected, and AML12 cells were treated with conditioned media for 4 h, after which fructose uptake in AML12 cells was measured by [3H]-fructose incorporation. (B) Fructose uptake in AML12 cells treated with fractions of the conditioned medium separated by a 3K filter using >3 kDa (proteins) or <3 kDa (metabolites) ( N = 3 samples per group, repeated in two independent experiments). (C) Fractions of MASLD media separated by fast protein liquid chromatography (FPLC) fractionation and activity in inducing fructose uptake in AML12 cells ( N = 2 samples per fraction for the fructose uptake assays). (D) Silver stain of all fractions of MASLD media separated by FPLC fractionation. (E) Number of total proteins and secreted proteins identified by unbiased proteomics. (F) Venn diagram showing the number of proteins that overlap between datasets. (G) Fold change of differentially expressed genes in male chow and MASLD mouse livers measured by single-cell RNA sequencing. (H) Angptl3 levels in hepatocyte-conditioned media from male mice measured by ELISA ( N = 3 samples, repeated in two independent experiments). (I) Fructose uptake measured by [3H]-fructose incorporation in AML12 cells after 24-h treatments with 100 nM recombinant mouse Angptl3 ( N = 3 samples per group, repeated in three independent experiments), Angptl4, or Tsk ( N = 3 samples per group). (J) Fructose uptake measured by [3H]-fructose incorporation in AML12 cells after 24-h treatments with 100 nM recombinant mouse Angptl3 or human insulin ( N = 3 samples per group, repeated in two independent experiments). (K) Circulating male mouse Angptl3 levels measured by ELISA ( N = 5 mice per group). (L) Weekly body weights of male mice fed fructose or glucose in drinking water ( N = 5 mice per group). (M) Water consumption in male mice fed fructose or glucose in drinking water ( N = 5 mice per group). (N) Weekly blood glucose of male mice fed fructose or glucose in drinking water ( N = 5 mice per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by two-tailed Student’s t test (I), one-way ANOVA (B, I, and J), or two-way ANOVA (K, L, M, and N).

Article Snippet: Angptl4 expression plasmid , OriGene , MR206437.

Techniques: Isolation, Cell Culture, Fast Protein Liquid Chromatography, Fractionation, Activity Assay, Silver Staining, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Recombinant, Two Tailed Test

Journal: Scientific Reports

Article Title: Mechanism of retinal angiogenesis induced by HIF-1α and HIF-2α under hyperoxic conditions

doi: 10.1038/s41598-025-20065-y

Figure Lengend Snippet: Hyperoxia leads to retinal astrocyte inactivation. ( A ) CCK-8 was used to assess the viability of astrocytes. ( B ) Flow cytometry was used to detect astrocyte apoptosis. ( C ) IF staining was used to visualize activated state astrocytes at 6 h, 12 h, and 24 h; scale bar: 100 μm; GFAP in green, PAX-2 in red. ( D , E ) ELISA was used to assess the levels of angiogenesis-related factors ANGPTL4(D) and VEGF(E) in astrocytes. ( F – J ) Western blot was used to visualize the protein expression of HIF-1α (F), HIF-2α (G), Laminin-β2 (H), dystrophin (I), and R-cadherin (J) in astrocytes at 24 h of culture. p < 0.05 was considered a significant difference, and n.s. means the difference was not significant. The data of RA and HO groups at 6 h, 12 h, and 24 h ( n = 3) were obtained through at least three independent repeated experiments.

Article Snippet: Cell supernatants were diluted 5-fold and assayed according to the instructions of the Mouse Angiopoietin-like Protein 4 (ANGPTL4) Kit (JL18884, Elabscience, China) and Mouse Vascular Endothelial Growth Factor A (VEGFA) Kit (E-MSEL-M0005, Elabscience, China).

Techniques: CCK-8 Assay, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Mechanism of retinal angiogenesis induced by HIF-1α and HIF-2α under hyperoxic conditions

doi: 10.1038/s41598-025-20065-y

Figure Lengend Snippet: Astrocyte inactivation leads to decreased function of RMECs. ( A ) Contact co-culture of astrocytes (green) with RMECs (red); scale bar: 500 μm. ( B , C ) ELISA was used to assess the levels of angiogenesis-related factors ANGPTL4(B) and VEGF(C) in the supernatants of contact co-cultured cells. ( D , E ) Western blot was used to visualize the protein expression of Laminin-β2(D) and integrin β1(E) in contact co-cultured astrocytes. ( F ) CCK-8 was used to assess the cell viability of RMECs in non-contact co-culture. G : Lumen formation assay was used to observe the tube-forming ability; scale bar: 300 μm. p < 0.05 was considered a significant difference, and n.s. means the difference was not significant. The data of RA and HO groups at 6 h, 12 h, and 24 h ( n = 3) were obtained through at least three independent repeated experiments.

Article Snippet: Cell supernatants were diluted 5-fold and assayed according to the instructions of the Mouse Angiopoietin-like Protein 4 (ANGPTL4) Kit (JL18884, Elabscience, China) and Mouse Vascular Endothelial Growth Factor A (VEGFA) Kit (E-MSEL-M0005, Elabscience, China).

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Expressing, CCK-8 Assay, Tube Formation Assay

Journal: Scientific Reports

Article Title: Mechanism of retinal angiogenesis induced by HIF-1α and HIF-2α under hyperoxic conditions

doi: 10.1038/s41598-025-20065-y

Figure Lengend Snippet: Overexpression of HIFs promotes the vasculature-directed capacity of astrocytes. ( A , B ) RT-qPCR was used to evaluate the overexpression of HIF-1α (A) and HIF-2α (B) at the mRNA level. ( C – F ) Western blot was used to evaluate the overexpression of HIF-1α (C) and HIF-2α (D) under room air and the overexpression of HIF-1α (E) and HIF-2α (F) under hyperoxia. ( G , H ) ELISA was used to assess the levels of ANGPTL4 (G) and VEGF (H). ( I – L ) Western blot was used to assess the protein expression of Laminin-β2 (I), integrin β1 (J), dystrophin (K), and R-cadherin (L). ( M ) IF staining was used to visualize HIF-1α, HIF-2α, Laminin-β2, and dystrophin in astrocytes after overexpression of HIFs under hyperoxia. p < 0.05 was considered a significant difference, and n.s. means the difference was not significant. The data of each group ( n = 3) was obtained through at least three independent repeated experiments.

Article Snippet: Cell supernatants were diluted 5-fold and assayed according to the instructions of the Mouse Angiopoietin-like Protein 4 (ANGPTL4) Kit (JL18884, Elabscience, China) and Mouse Vascular Endothelial Growth Factor A (VEGFA) Kit (E-MSEL-M0005, Elabscience, China).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Staining