Journal: Journal of biomedical science

Article Title: Neutrophil-derived reactive agents induce a transient SpeB negative phenotype in Streptococcus pyogenes.

doi: 10.1186/s12929-023-00947-x

Figure Lengend Snippet: Fig. 1 Reversible loss of SpeB is associated with tissue pathology and inflammation. A Distribution of SpeB+ and SpeB− GAS clones directly isolated from NSTI patient tissue biopsies (n = 23). B Percentage of SpeB+ and SpeB− GAS clones after the passage in THY media (p1, passage 1; p2, passage 2). Representative analysis of 2006 GAS patient isolate is shown. C–G Correlation analysis of bacterial load (left panel; n = 81 biopsies) or percentage of SpeB− clones (right panel; n = 23 biopsies) with the presence of HMGB1 (C), IL-8 (D), infiltrating neutrophils (E), and resistin (F) in patient biopsies. Correlation was determined using Spearman test. Semiquantitative acquired computerized image analyses (ACIA) of immuno-histochemical staining were performed as described in the methods section. The cell area was defined by the hematoxylin counterstaining, and the results are presented as percent positively stained area × mean intensity of positive staining

Article Snippet: The following antibodies were used for immunohistochemistry: anti-human HMGB1 (clone EPR3507; Abcam), anti-human IL-8 (clone NAP-1; Invitrogen), anti-human resistin (clone 184,305; R&D systems), and anti-human neutrophilelastase (clone NP57; DAKO).

Techniques: Clone Assay, Isolation, Staining

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Resistin Enhances Monocyte Chemoattractant Protein-1 Production in Human Synovial Fibroblasts and Facilitates Monocyte Migration.

doi: 10.33594/000000029

Figure Lengend Snippet: Fig. 5. Resistin promotes MCP-1-mediated monocyte migration by inhibiting miR-33a and miR-33b. (A&B) OASFs were incubated with resistin for 24 h and miRNA expression was examined by qPCR. (C) OASFs were transfected with the miR-33a and miR-33b mimics then incubated with resistin for 24 h; MCP-1 expression was measured by qPCR. (D) CM was applied to THP-1 cells and analyzed for migration activity. Results are expressed as the mean ± SEM. *p<0.05 as compared with the control group; #p<0.05 as compared with the resistin-treated group.

Article Snippet: KG Chen et al.: Resistin Enhances MCP-1-Mediated Monocyte Migration Materials and Methods Materials Recombinant human resistin was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, Incubation, Expressing, Transfection, Activity Assay, Control

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 1. Expression of Hresistin/RELMα during right ventricular dysfunction. A, Immunofluorescence images of RV tissue slices from monocrotaline (MCT)-treated rats and normal controls. Sections were stained with anti-RELMα (red), co-stained with anti-myosin (green), and counterstained with DAPI. Original magnification: ×200. The bottom panels show the outlined area from the 2-week post-MCT time point at higher magnification (×400). The arrowheads point to the myosin (green)-labeled cardiomyocytes positively stained for RELMα (red), whereas the arrows point to the myosin-negative RELMα-positive immune cells that infiltrated into the myocardial interstitium. Representative images are from 5 individual RV samples per group. B, Hresistin detection in human RV tissue (n=5 subjects per group). Confocal images show Hresistin (green signal) staining in the RV heart biopsy of non-PH control subjects and patients with scleroderma (SSc)-associated pulmonary arterial hypertension. Light micrograph of fluorescence images to show structure. Signals of light micrograph (showing tissue structure) and fluorescence images (of Hresistin and DAPI staining) are digitally merged and the boxed area is enlarged and displayed in the right panel. The arrowheads point to the Hresistin protein signals in cardiomyocytes whereas the arrows point to the Hresistin-positive staining in the infiltrating immune cell. C, Quantitative analysis of data in (A). Percentage of areas positive for RELMα in rat RV tissues was determined by the histogram tool with Adobe Photoshop software. Data are presented as mean±SEM (n=5 animals per group). *P<0.05 vs normal group. D, Quantitative real-time PCR analysis of RELMα genes in the right heart and left heart tissues of MCT-injected rats (n=5 animals per group; the dots in the graphs represent single individuals) with cardiac hypertrophy (2 weeks post-MCT) and failure (4 weeks post-MCT). E, Quantitative analysis of data in (B). The Hresistin-positive (+) cells were counted and expressed as numbers per high-power field (hpf). Data are presented as mean±SEM (n=5 subjects per group). *P<0.05. DAPI indicates 4’6-diamidino-2-phenyl-indole; Hresistin, human resistin; PCR, polymerase chain reaction; PH, pulmonary arterial hypertension; RELMα, resistin-like molecule-α; and RV, right ventricular.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Expressing, Immunofluorescence, Staining, Labeling, Control, Fluorescence, Software, Real-time Polymerase Chain Reaction, Injection, Polymerase Chain Reaction

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 2. Generation of cardiac-specific Hresistin transgenic mice. A and B, Images present the nucleotide sequence for the Hresistin (hRETN) construct (A) and a schematic representation of the αMHC-hRETN transgene structure (B). C, Immunofluorescence images of heart tissue samples from hRETN cardiac- overexpressing humanized mice and their corresponding littermate controls. Sections were stained with anti-myosin (green), co-stained with anti-FLAG (red), and counterstained with DAPI to validate the expression of the FLAG-tagged hRETN protein. Separate channels are displayed in middle and right panels, and digitally merged in left panels. Original magnification: ×200. Boxed areas are shown at higher magnification (×400) in the lower panels. Images are representative of 3 individual heart samples. D, Genotyping by PCR analysis of genomic DNA. Amplification of a 363-bp product encoding the myc-RETN epitope region of the transgene indicates that the humanized animals carry the knock-in hRETN gene in hearts. A 535-bp product was specifically amplified from animals carrying the tTA transgene. The 494-bp 18 S housekeeping gene served as a control. DAPI indicates 4’6-diamidino-2-phenyl-indole; MHC, myosin heavy chain; PCR, polymerase chain reaction; and tTA, tetracycline trans-activator.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Transgenic Assay, Sequencing, Construct, Immunofluorescence, Staining, Expressing, DNA Amplification, Knock-In, Amplification, Control, Polymerase Chain Reaction

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 3. Cardiac dysfunction and remodeling in humanized mice that overexpress MHC-hRETN. A, Pooled data of force-frequency (left panel) and intracellular Ca2+ transient-frequency (right panel) relationships in trabecular muscles from RV of wild-type and cardiac-specific MHC-hRETN-overexpressing mice (in force-frequency test: n=6 and 5 for control and overexpressing mice, respectively; in Ca2+ transient-frequency test: n=4 animals per group; the 2 assays used samples from different animals). *P<0.05, **P<0.01. B and C, Changes in phosphorylation of PKA (B) and AMPK (C) in cardiac tissue from cardiac-specific MHC-hRETN-overexpressing mice and control littermates were determined by western blotting. Left panels show representative immunoblots. Right panels show quantitative analysis of expression. Data are shown as mean±SEM (n = 6 animals per group). *P<0.05 vs control littermates. P- indicates phosphorylated protein; t- indicates total protein. D, Wheat germ agglutinin (WGA) cell boundary staining in the Hresistin-overexpressing myocytes. Left panels: heart tissue samples show cell nuclei (blue) and cell boundary (green). Magnification: ×400. Right panels: quantification of cell surface area based on histologic analysis of cardiomyocytes. Data represent means±SEM (n=5 animals per group). *P<0.05 vs littermate control group. E, Masson’s trichrome staining of heart tissue samples from MHC-Hresistin (RETN) humanized and littermate control groups. Magnification: ×200. Representative images (left panels) and quantitative analysis (right panels) were presented. Pooled data for quantification of fibrosis from 5 randomly selected histological fields at a magnification of ×400 on each slide. Data are shown as mean±SEM (n=5 animals per group). *P<0.05 vs control group. AMPK indicates AMPK-activated protein kinase; DAPI, 4’6-diamidino-2-phenyl-indole; hRETN, human resistin; MHC, myosin heavy chain; and PKA, protein kinase A.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Muscles, Control, Phospho-proteomics, Western Blot, Expressing, Staining

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 4. Hresistin activates HMGB1 signaling in the RV during RV r dysfunction. A, Immunofluorescence images of hypertrophic RV (RV-H, 2 weeks post-MCT induction) and failing RV (RV-F, 4 weeks post-MCT induction) from MCT-treated rats. Sections were stained with anti-HMGB1 antibody (green), costained with anti-myosin (red), and counterstained with DAPI (blue). Representative photographs of 4 individual animals per group. Original magnification: ×200. Boxed region in the RV-H group is shown at higher magnification to the right (×400). Further magnification (×1000) of the 2 framed areas are shown in the upper and lower panels to illustrate the HMGB1-positive, myosin-negative cells that infiltrated the myocardial interstitium. B and C, Quantitative analysis of data in A. Percentage of area positive for HMGB1 signal (HMGB1+) in rat right heart determined with Adobe Photoshop software (B) and the number (No.) of HMGB1-positive cells counted (C) on 5 randomly chosen RV fields in each animal at 200-fold magnification. Data are presented as mean±SEM (n=4 animals per group). *P<0.05, **P<0.01 vs normal (non-MCT-treated) rats. D, Immunofluorescence images of RV tissue from wild-type (WT) mice on post-hypoxia day 4 and from MHC-hRETN humanized mice. Some hypoxic (Hx) mice received daily intraperitoneal injections of the HMGB1-specific inhibitor ethyl pyruvate (EP, 50 mg/kg) for 4 days. Representative photographs of 4 individual animals per group. Original magnification: ×200. In the MHC-hRETN group, the corresponding co-staining for HMGB1 (green) with myosin (red) is presented in the lower panel, and the boxed region in it is shown at higher magnification (×400) on the left. E and F, Quantitative analysis of data in D. Percentage of area positive for HMGB1 signal in mouse right hearts was determined (E), and HMGB1- positive cells (per observed field) were counted (F). Data are presented as mean±SEM (n=5 animals per group for the control [normal WT] group and n=4 animals per group for the other 3 groups [Hx, Hx+EP, and MHC-hRETN]; the dots in the graphs represent single individuals). *P<0.05, **P<0.01 (increase) vs normal WT mice; †P<0.05 (decrease) vs the hypoxia (Hx)-only group. DAPI indicates 4’6-diamidino-2- phenyl-indole; EP, ethyl pyruvate; HMGB1, high mobility group box 1; hRETN, human resistin; hx, hypoxia; MCT, monocrotaline; MHC, myosin heavy chain; RV-F, failing right ventricle; and RV-H, hypertrophic right ventricle.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Immunofluorescence, Staining, Software, Control

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 5. Hresistin/HMGB1 signaling axis upregulates Ki67 expression in RV. A, Sections of RV tissues during hypertrophy or failure (as described in above panel A) were stained with anti-Ki67 antibody (green) and counterstained with DAPI (blue). Representative photographs of 4 individual animals per group. Magnification: ×200. B, Quantitative analysis of data from A. Ki67-positive (Ki67+) cells were counted on 5 randomly chosen fields of RV sections in each animal at ×200 magnification. Data are presented as mean±SEM (n=5 animals per group for the normal control group and n=4 animals per group for the groups of RV-H and RV-F; the dots in the graphs represent single individuals). *P<0.05 vs normal (non-MCT-treated) rats. C, Immunofluorescence images of RV tissue from WT mice exposed to 4 days of hypoxia with or without the HMGB1 inhibitor EP and from cardiac-specific MHC-hRETN-overexpressing mice. The boxed region in the fourth panel is shown at higher magnification (×400) on the right with costaining for Ki67 (green) and myosin (red). Representative photographs of 5 individual animals per group. Original magnification: ×200. D, Quantitative analysis of data in C. The Ki67-positive cells were counted (per observed field). Data are presented as mean±SEM (n=5 animals per group for the control [normal WT] group and n = 4 animals per group for the other 3 groups [Hx, Hx+EP, and MHC-hRETN]). *P<0.05, **P<0.01. E, In the RV tissues of the MHC-hRETN-overexpressing mice, Ki67 (red, middle panels) was further costained with the markers (green, left panels) of macrophages (F4/80), neutrophils (MPO), or fibroblasts (Vimentin). Images were merged in the right panels. The boxed areas were further enlarged in the far right panels showing the double positive cells. Representative photographs of 4 individual animals per group. Original magnification: ×400. DAPI indicates 4’6-diamidino-2-phenyl-indole; EP, ethyl pyruvate; HMGB1, high mobility group box 1; h-RETN, human resistin; Hx, hypoxia; MCT, monocrotaline; MHC, myosin heavy chain; MPO, myeloperoxidase; RV-F, failing right ventricle; RV-H, hypertrophic right ventricle; and WT, wild type.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Expressing, Staining, Control, Immunofluorescence

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 6. Anti-Hresistin human antibody ameliorates RV dysfunction in rats with PH. The anti-Hresistin antibody (Ab) or the isotype-matched control IgG1 (Con IgG) at 4 mg/kg were administered intraperitoneally twice a week in the hypoxia-induced PH rats. A and B, Echocardiographic analysis of right ventricular (RV) wall thickness and pulmonary artery blood velocity in Ab-treated hypoxic rats. RV wall thickness external diameter (RV-WTED) was measured as the distance from the free wall to the interventricular septum (millimeter) in the parasternal long-axis view using M-mode (A). Data are expressed as a percentage of the value of normoxic control mice. The anti-Hresistin Ab treatment also lengthened pulmonary artery acceleration time (PAT). Results of pulsed wave Doppler measurement of PAT are shown in B. PAT values were normalized by pulmonary ejection time (PET). Data are expressed as means±SEM (n=6 animals per group). *P<0.05, **P<0.01 vs hypoxia (no Ab) group. Representative echocardiographic images are shown in the right panels. C, Immunoprecipitation analysis of the binding of rat RELMα to the human therapeutic Ab targeting Hresistin. The protein-Ab binding was detected by western blotting with the anti-Hresistin antibody from R&D (AF1359). Recombinant rat RELMα protein was loaded as the positive control. D and E, Analysis of RV hypertrophy and hemodynamics in the hypoxia (Hx)-induced rat PH model. We measured the RV systolic pressure (RVSP) (D) and Fulton index (ratio of RV weight/ LV+S weight) (E). Data are presented as means±SEM (normal no Ab: n=6, normal con Ab: n=6, hypoxia no Ab: n=5, hypoxia con Ab: n=6, hypoxia Ab: n=5; n refers to the number of animals per group). **P<0.01 vs Hx-treated group without Ab treatment. F, Results of treatment with the human antibody targeting Hresistin to improve the survival rate among rats with monocrotaline-induced PH. *P<0.05 by log rank test; n=6 animals in each group. LV+S indicates left ventricle plus septum; MCT, monocrotaline; PH, pulmonary arterial hypertension; and RELMα, resistin-like molecule-α.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Control, Immunoprecipitation, Binding Assay, Western Blot, Recombinant, Positive Control

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 7. Hresistin/HMGB1 signaling axis induction of cardiac dysfunction and hypertrophy. A, Representative immunoblots of the protein levels of RELMα, HMGB1, and the hypertrophy markers for atrial natriuretic peptide (ANP) and myosin heavy chain-β (β-MHC) in RV showing anti-Hresistin Ab inhibition of the hypoxia-induced expression of HMGB1 and RV hypertrophy markers in the RV of PH rats in vivo. B, Quantitative analysis of data in A. n=4 rats per group, one-way ANOVA with Tukey post hoc analysis for multiple group comparisons. C, Immunoblots showing anti-Hresistin Ab prevention of the Hresistin- induced expression of HMGB1 and RV hypertrophy markers in the neonatal rat cardiomyocytes (NRCMs) in vitro. The primary cultured NRVMs were pretreated with 3 μg/mL Con IgG or the anti-Hresistin Ab followed by transduction of MOI-300 GFP-tagged adeno- associated virus (AAV) expressing the Hresistin (hRETN). Empty AAV vector (null) served as the negative control. Representative immunoblots are shown (n=4 per group). D, Quantitative analysis of data in C. Data are expressed as means±SEM (n=4 per group). *P<0.05, **P<0.01, ***P<0.001. E through G, Representative and quantitative WB images and quantitative analysis of the HMGB1 inhibitor ethyl pyruvate (EP) attenuation of the Hresistin-induced cardiac hypertrophy in NRCMs in vitro. The primary NRVMs were pretreated with 5 μmol/L EP followed by MOI-300 AAV-hRETN transduction. Representative WB images (E) and quantitative analysis (F) of ANP and β-MHC protein are displayed (n=4 per group). Quantification of the cell surface area of NRCMs is shown in G (n=13 per group). *P<0.05, **P<0.01. Ab indicates antibody; GFP, green fluorescent protein; HMGB1, high mobility group box 1; Hresistin, human resistin; Hx, hypoxia; NRVM, neonatal rat cardiomyocyte; PH, pulmonary arterial hypertension; RELMα, resistin-like molecule-α; RV, right ventricle; and WB, western blot.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques: Western Blot, Inhibition, Expressing, In Vivo, In Vitro, Cell Culture, Transduction, Virus, Plasmid Preparation, Negative Control

Journal: Journal of the American Heart Association

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension

doi: 10.1161/jaha.122.027621

Figure Lengend Snippet: Figure 8. Schematic illustration of Hresistin-induced cardiac inflammation and dysfunction. During pulmonary arterial hypertension (PH) development, Hresistin activates the damage-associated molecular pattern (DAMP), signaling triggering of inflammation in the right ventricle (RV) and contributing to RV dysfunction pathogenesis. Targeting the Hresistin signaling cascade may constitute a novel therapeutic approach to RV dysfunction and other related cardiac diseases in humans. HMGB1 indicates high mobility group box 1; Hresistin, human resistin; LV, left ventricle; and RELMα, resistin-like molecule-α.

Article Snippet: In the monocrotaline model, rat RV sections were treated with anti- RELMα (AF1523, R&D Systems) and anti- myosin (ab185967, Abcam) antibodies, whereas human RV tissue from SSc- PH patients were treated with anti- Hresistin antibody (AF1359, R&D Systems) for immune staining.

Techniques:

Journal: Irish Journal of Medical Science

Article Title: Cardioprotective properties of leptin in patients with excessive body mass

doi: 10.1007/s11845-020-02211-9

Figure Lengend Snippet: Evaluation of biochemical parameters among the groups

Article Snippet: The Enzyme Linked-Immunosorbent Assay (ELISA) tests were conducted for quantitative determination of resistin (BioVendor, Czech Republic), adiponectin (BioVendor, Czech Republic), leptin (BioVendor, Czech Republic), IL-6 (Gen-Probe, France), ADMA (Immunodiagnostic, Bensheim, Germany), and PIIINP (Cloud-Clone Corp, China) in human serum.

Techniques:

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 1. Immunohistochemical staining of aneurysms with antibodies to resistin and CD68. The various types of human vessels were stained with antibodies against resistin or CD68, the latter of which is a marker of monocytes/macrophages. Mayer’s hematoxylin was used for counterstaining. The left upper region of each picture indicates the endothelial lining. Resistin-positive areas are seen in atherosclerotic lesions as dark staining beneath the endothelial lining (C and D). Resistin-positive areas are distributed along inflammatory cells and stained positively for CD68 (G, H). No immunoreactive area was observed for either of the two antibodies in normal vessels (A, B, E, F). Staining without primary Abs was performed as a negative control (I, J, K, L).

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Immunohistochemical staining, Staining, Marker, Negative Control

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 3. Resistin mRNA expression measured by real-time PCR. (A) Resistin mRNA expression in aortic aneurysms. Total RNA was purified from aneurysm centers (n =4) and veins (n =3). Real-time quantitative PCR revealed the overexpression of resistin in aneurysm centers. The y axis represents resistin expression normalized against GAPDH expression. The data illustrated on the graph represent meansTSEM of 2deltadeltaCt of the samples. (B) Resistin mRNA expression in vascular cells. Isolated human VSMCs and commercial HUVECs were incubated with 10 ng/mL of TNF-a for 12 h. Human monocytes were isolated and incubated for 36 h, and total RNA was purified and resistin mRNA levels were measured by real-time RT-PCR, normalized with respect to the GAPDH mRNA levels. The data illustrated on the graph represent meansTSEM of 2deltadeltaCt of three different experiments.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Purification, Over Expression, Isolation, Incubation, Quantitative RT-PCR

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 2. Immunofluorescent double staining of aneurysms with antibodies to resistin and vascular cell markers. Green indicates vascular cell proteins, red resistin, and blue DAPI-stained cellular nuclei. The upper region in each picture indicates endothelial lining. Resistin protein was found to co-localize with CD68 (A, B). Smooth muscle (SM) actin was positive throughout the vessel thickness (D), whereas CD31 was not detected (G). The distribution of resistin (E, H) was not concordant with either D or G. A, B and C were magnified (J¨L) and this showed that resistin was present both intracellularly and extracellularly. The merged picture (M) shows yellow rings composed of red resistin-positive areas and green transmembrane CD68-positive areas (arrows).

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Double Staining, Staining

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 4. Induction of PAI-1 in HUVECs after resistin treatment. HUVECs were incubated with 10¨100 ng/mL of resistin for 4¨24 h or with 10 ng/mL of TNF- a for 12 h as a positive control. (A) PAI-1 protein levels in culture media were measured by ELISA and are presented as fold changes versus the negative control. (B) PAI-1 mRNA expression was analyzed by Northern blotting, and normalized against GAPDH mRNA. Results are presented as fold changes versus the negative control. The data illustrated on the graph represent the meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Incubation, Positive Control, Enzyme-linked Immunosorbent Assay, Negative Control, Expressing, Northern Blot

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 6. Induction of VSMC migration by resistin. VSMCs in serum-free media were incubated with 10¨100 ng/mL resistin for 24 h or with 10% FBS for 24 h as a positive control. Cell migration was detected by scratched wound assays and results are presented as fold changes versus the negative control. The data illustrated on the graph represent meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Migration, Incubation, Positive Control, Negative Control

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 5. Induction of ET-1 in HUVECs after resistin treatment. HUVECs were incubated with 10¨100 ng/mL of resistin for 4¨24 h or with 10 ng/mL of TNF-a for 12 h as a positive control. ET-1 mRNA levels were analyzed by Northern blotting and normalized against GAPDH mRNA, and are presented as fold changes versus the negative control. The data illustrated on the graph represent the meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Incubation, Positive Control, Northern Blot, Negative Control