polyclonal rabbit anti cd71  (Cell Signaling Technology Inc)

Journal: Frontiers in Cardiovascular Medicine

Article Title: Exosomes derived from pericardial adipose tissues attenuate cardiac remodeling following myocardial infarction by Adipsin-regulated iron homeostasis

doi: 10.3389/fcvm.2022.1003282

Figure Lengend Snippet: Adipsin-enriched pericardial-AT exosomes regulate iron homeostasis and alleviate lipid oxidative stress in vivo . (A,B) Non-heme iron levels in the serum and the peri-infarct region of the cardiac tissue were assessed in MI mouse after treatment with Adipsin-transgenic or non-transgenic exosomes. Each group n = 6, * p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-NTg mice, I p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-Tg mice, § p < 0.05 vs. MI + Pericardial-AT exosomes from Adipsin-NTg mice. Data were mean ± SEM, and one-way ANOVA was used for statistical analysis. (C) The levels of malondialdehyde (MDA) in cardiac tissue were measured in the peri-infarct area. Each group n = 6, * p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-NTg mice, I p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-Tg mice, § p < 0.05 vs. MI + Pericardial-AT exosomes from Adipsin-NTg mice. Data were mean ± SEM, and one-way ANOVA was used for statistical analysis. (D) Perls’ Prussian Blue-stained heart slices. (E–I) TFRC, FTH, COX2, and GPX4 proteins were measured by Western blots. Each group n = 6, * p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-NTg, I p < 0.05 vs. Sham + Pericardial-AT exosomes from Adipsin-Tg, § p < 0.05 vs. MI + Pericardial-AT exosomes from Adipsin-NTg. Data were mean ± SEM, and one-way ANOVA was used for statistical analysis.

Article Snippet: Antibodies used included anti-Adipsin rabbit monoclonal antibody (Abcam, ab213177, 1/1,000), anti-transferrin rabbit monoclonal antibody (Abcam, ab109503, 1/8,000), HRP anti-beta actin mouse monoclonal antibody (Abcam, ab20272, 1/5,000), anti-TFRC rabbit polyclonal antibody (Proteintech, 10084-2-AP, 1/2,000), anti-ferritin heavy chain rabbit monoclonal antibody (Abcam, ab183781, 1/1,000), anti-COX2 rabbit polyclonal antibody (ABclonal, A1253, 1/3,000), anti-GPX4 rabbit monoclonal antibody (ABclonal, A11243, 1/1,000), anti-GAPDH rabbit polyclonal antibody (Proteintech, 10494-1-AP, 1/8,000).

Techniques: In Vivo, Transgenic Assay, Staining, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Exosomes derived from pericardial adipose tissues attenuate cardiac remodeling following myocardial infarction by Adipsin-regulated iron homeostasis

doi: 10.3389/fcvm.2022.1003282

Figure Lengend Snippet: Irp2 is essential to Adipsin-offered cardioprotection. (A) Flowchart of the study design. Myocardial infarction surgery was performed 14 days after injection of AAV9 (Scramble or IRP2-shRNA) at the area of the myocardium. Following that, MI mice were injected four times with exosomes isolated from Adipsin-Tg or NTg mice’s pericardial adipose tissues. Cardiac function was assessed 28 days after MI surgery. Each group n = 6. (B,C) Representative echocardiographic images and left ventricular ejection fraction (LVEF) analysis. Each group n = 6. * p < 0.05 vs. AAV9-Scramble + Exo (NTg) group. (D) Non-heme iron levels were measured in the peri-infarct region of heart. Each group n = 6. * p < 0.05 vs. AAV9-Scramble + Exo (NTg) group. (E,F) Quantification of fibrotic area and representative images of Masson’s trichrome staining of the transverse planes post myocardial infarction. Each group n = 6. * p < 0.05 vs. AAV9-Scramble + Exo (NTg) group. (G) Malondialdehyde (MDA) levels in the peri-infarct region of heart. Each group n = 6. * p < 0.05 vs. AAV9-Scramble + Exo (NTg) group. (H–L) TFRC, FTH, COX2, and GPX4 proteins were measured by Western blots. Each group n = 6. * p < 0.05 vs. AAV9-Scramble + Exo (NTg) group. Data were mean ± SEM, and one-way ANOVA was used for statistical analysis.

Article Snippet: Antibodies used included anti-Adipsin rabbit monoclonal antibody (Abcam, ab213177, 1/1,000), anti-transferrin rabbit monoclonal antibody (Abcam, ab109503, 1/8,000), HRP anti-beta actin mouse monoclonal antibody (Abcam, ab20272, 1/5,000), anti-TFRC rabbit polyclonal antibody (Proteintech, 10084-2-AP, 1/2,000), anti-ferritin heavy chain rabbit monoclonal antibody (Abcam, ab183781, 1/1,000), anti-COX2 rabbit polyclonal antibody (ABclonal, A1253, 1/3,000), anti-GPX4 rabbit monoclonal antibody (ABclonal, A11243, 1/1,000), anti-GAPDH rabbit polyclonal antibody (Proteintech, 10494-1-AP, 1/8,000).

Techniques: Injection, shRNA, Isolation, Staining, Western Blot

Journal: eLife

Article Title: Aging is associated with increased brain iron through cortex-derived hepcidin expression

doi: 10.7554/eLife.73456

Figure Lengend Snippet: mRNA levels of proteins involved in iron regulation in the liver ( A ) and brain ( B ) in young (4 months old) and old (22 months old) mice (n=10/group). Means and SEM are indicated as horizontal and vertical bars, respectively. An undetected measurement (n=1) in the brain Fech and undetected measurements (n=2) and an outlier (n=1) in the brain Hamp1 were excluded. Tfr1 = transferrin receptor 1, Tfr2 = transferrin receptor 2, Dmt1 = divalent metal transporter 1, Fpn1 = ferroportin 1, Pcbp1 = Poly(RC) Binding Protein 1, Steap3 = Metalloreductase Six-Transmembrane Epithelial Antigen Of Prostate 3, Sec15l1 = exocyst complex component 6, Mfrn2 = mitoferrin 2, Abcb7 = ATP-binding cassette sub-family B member 7, Abcb8 = ATP-binding cassette sub-family B member 8, Alr1 = augmenter of liver regeneration, Cp = ceruloplasmin, Fxn = frataxin, Ttp = tristetraprolin, Lias = tristetraprolin, Hmox1 = heme oxygenase 1, Hmox2 = heme oxygenase 2, Alas1 = 5′-aminolevulinate synthase 1, Alas2 = 5′-aminolevulinate synthase 2, Fech = ferrochelatase, Alad = aminolevulinate dehydratase, Pbgd = porphobilinogen deaminase, Uros = uroporphyrinogen III synthase, Urod = uroporphyrinogen decarboxylase, Ppox = protoporphyrinogen oxidase, Abcb10 = ATP-binding cassette, sub-family B member 10, Hamp1 = hepcidin1, Bmp6 = bone morphogenetic protein 6, Hfe = homeostatic iron regulator, Ftmt = mitochondrial ferritin. ( C ) Representative immunoblot for hepcidin1 in the brain (n=6). ( D ) Summary of densitometry analysis of panel ( C ). ( E ) Representative immunohistochemistry of hepcidin1 (Red = anti-hepcidin1, blue = DAPI) in the brain frontal cortex of young and aged mice. Scale bar=200 µm. * p<0.05. Figure 2—source data 1. Full-length images of immunoblots shown in . Figure 2—source data 2. qRT-PCR liver – Original data of qRT-PCR in the liver shown in . Figure 2—source data 3. qRT-PCR brain – Original data of qRT-PCR in the brain shown in . Figure 2—source data 4. Densitometry – Original data of densitometry quantifications of immunoblots shown in .

Article Snippet: Antibody , Rabbit polyclonal anti-TFR1 antibody , ProteinTech , 100-84-2-AP , WB (1:1000).

Techniques: Binding Assay, Western Blot, Immunohistochemistry, Quantitative RT-PCR

Journal: eLife

Article Title: Aging is associated with increased brain iron through cortex-derived hepcidin expression

doi: 10.7554/eLife.73456

Figure Lengend Snippet: ( A ) Immunoblots of iron transporting proteins TfR1 and FPN1 in the brain cortex of young and aged mice (n=3). ( B ) Summary of densitometric analysis of panel ( A ). ( C ) Poly-ubiquitination levels of FPN1, as assessed by immunoprecipitation, in the brain cortex of young and aged mice (n=3). ( D ) Summary of the densitometric analysis of panel ( C ). ( E ) Fe-S cluster containing aconitase enzyme activity, a marker of cellular oxidative stress, in the brain cortex of young and aged mice (n=4). c-aconitase = cytosolic aconitase (ACO1), m-aconitase = mitochondrial aconitase (ACO2). * p<0.05. Figure 3—source data 1. Full length images of immunoblots shown in . Figure 3—source data 2. Densitometry – Original data of densitometry quantifications of immunoblots shown in . Figure 3—source data 3. Aconitase assay – Original data of aconitase enzyme activities shown in .

Article Snippet: Antibody , Rabbit polyclonal anti-TFR1 antibody , ProteinTech , 100-84-2-AP , WB (1:1000).

Techniques: Western Blot, Ubiquitin Proteomics, Immunoprecipitation, Activity Assay, Marker

Journal: eLife

Article Title: Aging is associated with increased brain iron through cortex-derived hepcidin expression

doi: 10.7554/eLife.73456

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-TFR1 antibody , ProteinTech , 100-84-2-AP , WB (1:1000).

Techniques: Protease Inhibitor, cDNA Synthesis, SYBR Green Assay, Immunoprecipitation, Bicinchoninic Acid Protein Assay, Isolation, Activity Assay, Sequencing, Software

Journal: eLife

Article Title: Aging is associated with increased brain iron through cortex-derived hepcidin expression

doi: 10.7554/eLife.73456

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-TFR1 antibody , ProteinTech , 100-84-2-AP , WB (1:1000).

Techniques: