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af1523 (R&D Systems)

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R&D Systems af1523
Af1523, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/af1523/Mouse+FIZZ1%2FRELM+alpha+Antibody/pmc13068211-75-17-15
Average 93 stars, based on 6 article reviews

af1523 - by Bioz Stars, 2026-09

93/100 stars

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Electrophoresis:

Article Title: Human resistin is critical to activation of the NLRP3 inflammasome in macrophages
Article Snippet: The supernatants were mixed in sodium dodecyl sulfate (SDS) 5× sample loading buffer (NuPAGE, Invitrogen) at 99°C for 10 minutes and then subjected to SDS-polyacrylamide gel electrophoresis (PAGE). .. After electrophoresis, proteins were transferred onto polyvinylidene difluoride membranes and immunoblotted with antibodies to RELMα (R&D Systems, AF1523), β-actin (Cell Signaling Technology, 3700), BTK (Sigma-Aldrich, SAB4502936), HMGB1 (Abcam, ab79823), NLRP3 (Abcam, ab214185), pro- and cleaved caspase-1 (Abcam, ab207802), pro- and cleaved IL-18 (Invitrogen, PA5-6082), and pro- and cleaved IL-1β (Abcam, ab254360) overnight at 4°C and then probed with horseradish peroxidase–conjugated secondary antibodies (Bio-Rad) for 2 hours. .. Protein bands were visualized by chemiluminescence (ECL, Amersham Pharmacia Biotech).

Article Title: Human Resistin Induces Cardiac Dysfunction in Pulmonary Hypertension
Article Snippet: The supernatants were mixed in SDS sample loading buffer (NuPAGE, Invitrogen) at 99 °C for 10 minutes and then subjected to SDS- PAGE. .. After electrophoresis, proteins were transferred onto polyvinylidene difluoride membranes and immunoblotted with antibodies to RELMα (AF1523, R&D Systems), total PKA C (4782, Cell Signaling), phospho- PKA C (5661, Cell Signaling), total AMPK (Ab131512, Abcam), phospho- AMPK (50 081, Cell Signaling), HMGB1 (ab18256, Abcam), Ki67 (ab16667, Abcam), or GAPDH (G8795, Sigma) overnight at 4 °C, and then probed with horseradish peroxidase– conjugated secondary antibodies (Bio- Rad) for 2 hours. .. Protein bands were visualized by chemiluminescence (ECL; Amersham Pharmacia Biotech, Arlington Heights, IL).

Article Title: Human resistin is critical to activation of the NLRP3 inflammasome in macrophages
Article Snippet: The supernatants were mixed in sodium dodecyl sulfate (SDS) 5 × sample loading buffer (NuPAGE, Invitrogen) at 99°C for 10 minutes and then subjected to SDS-polyacrylamide gel electrophoresis (PAGE). .. After electrophoresis, proteins were transferred onto polyvinylidene difluoride membranes and immunoblotted with antibodies to RELMα (R&D Systems, AF1523), β-actin (Cell Signaling Technology, 3700), BTK (Sigma-Aldrich, SAB4502936), HMGB1 (Abcam, ab79823), NLRP3 (Abcam, ab214185), pro- and cleaved caspase-1 (Abcam, ab207802), pro- and cleaved IL-18 (Invitrogen, PA5−6082), and pro- and cleaved IL-1β (Abcam, ab254360) overnight at 4°C and then probed with horseradish peroxidase–conjugated secondary antibodies (Bio-Rad) for 2 hours. .. Protein bands were visualized by chemiluminescence (ECL, Amersham Pharmacia Biotech).

Western Blot:

Article Title: Therapeutic Benefit of Melatonin in Choroidal Neovascularization During Aging Through the Regulation of Senescent Macrophage/Microglia Polarization
Article Snippet: .. CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) .. Number Dilution CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) Arg-1, arginase-1; Fizz-1, resistin-like-α; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Iba1, ionized calcium‐binding adaptor molecule 1; IF, immunofluorescence; IL-10R, interleukin-10 receptor; STAT3, signal transducer and activator of transcription 3; WB, western blot; Ym-1, chitinase 3-like 3.

Article Title: Therapeutic Benefit of Melatonin in Choroidal Neovascularization During Aging Through the Regulation of Senescent Macrophage/Microglia Polarization
Article Snippet: CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) .. Number Dilution CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) Arg-1, arginase-1; Fizz-1, resistin-like-α; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Iba1, ionized calcium‐binding adaptor molecule 1; IF, immunofluorescence; IL-10R, interleukin-10 receptor; STAT3, signal transducer and activator of transcription 3; WB, western blot; Ym-1, chitinase 3-like 3. ..

Immunofluorescence:

Article Title: Therapeutic Benefit of Melatonin in Choroidal Neovascularization During Aging Through the Regulation of Senescent Macrophage/Microglia Polarization
Article Snippet: CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) .. Number Dilution CD31 Millipore (Darmstadt, Germany) MAB1398Z 1:200 (IF) Arg-1 Cell Signaling (Beverly, CA, USA) 93668S 1:1000 (WB) YM-1 Abcam (Cambridge, UK) ab93034 1:500 (WB) Fizz-1 R&D Systems (Minneapolis, MN) AF1523 1:1000 (WB) CD206 R&D Systems (Minneapolis, MN) AF2535 1:100 (IF) Iba1 Wako Pure Chemical Industries (Osaka, Japan) 019-19741 1:100 (IF) p16 Abcam (Cambridge, UK) ab189034 1:100 (IF) 1:1000 (WB) p21 Abcam (Cambridge, UK) ab109199 1:1000 (WB) p53 Abcam (Cambridge, UK) ab131442 1:500 (WB) IL-10R1 Abcam (Cambridge, UK) ab225820 1:1000 (WB) IL-10R2 R&D Systems (Minneapolis, MN) AF5368 1:1000 (WB) p-STAT3 Cell Signaling (Beverly, CA, USA) 4113S 1:100 (IF) 1:1000 (WB) t-STAT3 Cell Signaling (Beverly, CA, USA) 9139S 1:1000 (WB) GAPDH Servicebio Technology Co. (Wuhan, China) GB11002 1:2000 (WB) Arg-1, arginase-1; Fizz-1, resistin-like-α; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Iba1, ionized calcium‐binding adaptor molecule 1; IF, immunofluorescence; IL-10R, interleukin-10 receptor; STAT3, signal transducer and activator of transcription 3; WB, western blot; Ym-1, chitinase 3-like 3. ..

Image Search Results

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.

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

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.

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.

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

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.

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.

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

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af1523 (R&D Systems)

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