Journal: Biomedicines

Article Title: Irisin—A Pancreatic Islet Hormone

doi: 10.3390/biomedicines10020258

Figure Lengend Snippet: Confocal image of a single islet of Langerhans from ( A – C ) mouse, ( D – F ) rat and ( G – I ) human split into fibronectin type III domain-containing protein 5 (FNDC5, green), insulin (red) and a combined picture. Scale bar = 100 μm.

Article Snippet: For blocking, 3% donkey serum/PBS was used for 30 min, followed by overnight incubation at 4 °C with primary antibodies for FNDC5 (Rabbit polyclonal antibody bs-8486R, Bioss, Woburn, MA, USA) and insulin (Cat No. 20-IP30, Fitzgerald, Acton, MA, USA) or glucagon (Biotin conjugated antibody, Cat No. 13-9743-82, eBioscience™, ThermoFisher scientific, Waltham, MA, USA).

Techniques:

Journal: Biomedicines

Article Title: Irisin—A Pancreatic Islet Hormone

doi: 10.3390/biomedicines10020258

Figure Lengend Snippet: Confocal image of a single islet of Langerhans from ( A – C ) mouse, ( D – F ) rat and ( G – I ) human split into FNDC5 (green), glucagon (red) and a combined picture. Scale bar = 100 μm.

Article Snippet: For blocking, 3% donkey serum/PBS was used for 30 min, followed by overnight incubation at 4 °C with primary antibodies for FNDC5 (Rabbit polyclonal antibody bs-8486R, Bioss, Woburn, MA, USA) and insulin (Cat No. 20-IP30, Fitzgerald, Acton, MA, USA) or glucagon (Biotin conjugated antibody, Cat No. 13-9743-82, eBioscience™, ThermoFisher scientific, Waltham, MA, USA).

Techniques:

Journal: Biomedicines

Article Title: Irisin—A Pancreatic Islet Hormone

doi: 10.3390/biomedicines10020258

Figure Lengend Snippet: Relative expression of the gene fibronectin type III domain-containing protein 5 (FNDC5) in rat and human islets compared to skeletal muscle, calculated using the housekeeping genes glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and Ribosomal Protein S7 (RPS7) separately. Each value represents one animal or human donor.

Article Snippet: For blocking, 3% donkey serum/PBS was used for 30 min, followed by overnight incubation at 4 °C with primary antibodies for FNDC5 (Rabbit polyclonal antibody bs-8486R, Bioss, Woburn, MA, USA) and insulin (Cat No. 20-IP30, Fitzgerald, Acton, MA, USA) or glucagon (Biotin conjugated antibody, Cat No. 13-9743-82, eBioscience™, ThermoFisher scientific, Waltham, MA, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Comparison of FNDC5/Ir (( A )—IRS score 8) expression with E-cadherin (( B )—IRS score 12), N-cadherin (( C )—IRS score 8), SNAIL (( D )—IRS score 3 and % of nuclear expression 4, ( E )—IRS score 12 and % of nuclear expression 4), SLUG (( F )—IRS score 8 and % of nuclear expression 3) and TWIST (( G )—IRS score 4 and % of nuclear expression 2, ( H )—IRS score 4 and % of nuclear expression 2) using immunohistochemistry (IHC) (positive reactions—brown cell cytoplasm) in breast cancer (BC), magnification ×200.

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Correlation of FNDC5/Ir expression level with E-cadherin ( A ) and N-cadherin ( B ), cytoplasmic ( C ) and nuclear ( D ) SNAIL expression levels, cytoplasmic ( E ) and nuclear ( F ) SLUG expression levels, cytoplasmic ( G ) and nuclear ( H ) TWIST expression levels in breast cancer (BC) (sample size n = 541).

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Comparison of mRNA FNDC5 expression levels detected by RT-PCR ( A ) and FNDC5/Ir levels ( B ) in the normal breast cell line (Me16c) and different types of BC cell lines (MCF-7, MDA-MB-231, MDA-MB-468) * p < 0.05 ** p < 0.01 *** p < 0.001.

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Comparison of FNDC5/Ir expression by confocal microscopy in the normal breast cell line (Me16c) and different BC cell lines (MCF-7, MDA-MB-231, MDA-MB-468).

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing, Confocal Microscopy

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Immunolocalization of Ir in transmission electron microscopy. Ultrathin section examination of human adenocarcinoma BC cells (from tumors, ( A – D ) and MDA-MB 468 cell line, ( E , F )). The specific primary antibody against FNDC5/Ir was applied. Next, the ultrathin sections were labeled with the secondary antibody conjugated with the 20 nm-colloidal gold nanoparticles, which shows the antigen distribution in the cells. Arrows indicate positive gold nanoparticles. A strong reaction was detected in the cytoplasm of cancer cells, in the mitochondria, and at the border of the cell membranes of neighboring cells. Note the localization of Ir near the specific microvilli–like structure ( E ) and at the cytoplasmatic processes of BC cells ( F ). Brief double staining with UranyLess solution and lead citrate (3%). IDC—invasive ductal carcinoma at different grades of malignancy, N—nucleus, Mi—mitochondrion.

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Transmission Assay, Electron Microscopy, Labeling, Double Staining

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Immunolocalization of Ir in transmission electron microscopy. Ultrathin section examination of human adenocarcinoma BC cells from the breast tumor microenvironment (stroma). All electronograms show invasive ductal carcinoma G2. The specific primary antibody against FNDC5/Ir was applied as previously described, followed by applying the species-specific secondary antibody conjugated with 20 nm-colloidal gold nanoparticles. Arrows indicate positive gold nanoparticles. Strong immunogold reaction was detected in the extracellular matrix and cancer-associated fibroblasts (in the cytoplasm and at the border of the cell membrane). Brief double staining with UranyLess solution and lead citrate (3%). N—nucleus, Gs—ground substance of the extracellular matrix ( A ), Cf—collagen fibers, RER—rough endoplasmic reticulum in the fibroblast ( B ). IF—intermediate filaments ( C ), Mi—mitochondrion ( D ).

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Transmission Assay, Electron Microscopy, Double Staining

Journal: Heliyon

Article Title: Astragaloside IV inhibits angiotensin II-induced atrial fibrosis and atrial fibrillation by SIRT1/PGC-1α/FNDC5 pathway

doi: 10.1016/j.heliyon.2024.e30984

Figure Lengend Snippet: AS-IV suppressed the expression of fibrotic genes and SIRT1, PGC-1α and FNDC5 proteins in Ang II-treated mice. (A) The mRNA expression of α-SMA, Collagen I and Collagen III. (B) Representative bands of SIRT1, PGC-1α and FNDC5 by Western blot. (C) Quantification of SIRT1, PGC-1α and FNDC5 proteins. Protein expression was normalized to the expression of GAPDH. (D) Serum irisin concentration in the mice of four groups. Data are presented as mean ± SD (n = 8 in each group). ***p < 0.001 vs. control group; ###p < 0.001 vs. Ang II group. Abbreviation: SIRT1, sirtuin-1; PGC-1α, peroxisome proliferator-activated receptor gamma co-activator 1-alpha; FNDC5, fibronectin type III domain containing 5.

Article Snippet: The following antibodies were incubated on the membrane for one night at 4 °C: FNDC5 (1:400, ab131390, rabbit polyclonal, Abcam), PGC-1α (1:500, ab54481, rabbit polyclonal, Abcam), and GAPDH (1:1000, ab9485, rabbit polyclonal, Abcam).

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Heliyon

Article Title: Astragaloside IV inhibits angiotensin II-induced atrial fibrosis and atrial fibrillation by SIRT1/PGC-1α/FNDC5 pathway

doi: 10.1016/j.heliyon.2024.e30984

Figure Lengend Snippet: Schematic illustration of the protective effects of AS-IV against Ang II-induced atrial fibrosis and AF via upregulation of the SIRT1/PGC-1α/FNDC5 pathway.

Article Snippet: The following antibodies were incubated on the membrane for one night at 4 °C: FNDC5 (1:400, ab131390, rabbit polyclonal, Abcam), PGC-1α (1:500, ab54481, rabbit polyclonal, Abcam), and GAPDH (1:1000, ab9485, rabbit polyclonal, Abcam).

Techniques:

Journal: Balkan Medical Journal

Article Title: The Effects of Irisin on Nω-Nitro-L-arginine Methyl Ester Hydrochloride-Induced Hypertension in Rats

doi: 10.4274/balkanmedj.galenos.2019.2019.5.113

Figure Lengend Snippet: a-d. Glomerular immunostaining for endothelial nitric oxide synthase in different groups (x200). In all groups, widespread strong staining of glomerular capillaries (red arrow) and weak staining of proximal tubules were observed. When the Hypertension and Hypertension + irisin groups are compared, the Hypertension + irisin group was observed to have a significantly reduced level of staining (p<0.01). Control group (a); Irisin group (b); Hypertension group (c); Hypertension + irisin group (d).

Article Snippet: The immunohistochemical antibody [irisin (Bioss FNDC5 BS-4886-R), inducible NOS (Spring REF E3744), and endothelial NOS (Neomarkers RB-9279-P)] panel used for the diagnosis and differential diagnosis varies according to the cases and cytokeratin.

Techniques: Immunostaining, Staining

Journal: Balkan Medical Journal

Article Title: The Effects of Irisin on Nω-Nitro-L-arginine Methyl Ester Hydrochloride-Induced Hypertension in Rats

doi: 10.4274/balkanmedj.galenos.2019.2019.5.113

Figure Lengend Snippet: a-d. Glomerular immunostaining for inducible nitric oxide synthase in different groups (x200). While the control and B groups were observed to have focal and mild staining [glomeruli (red arrow), tubules (blue arrow)], there was widespread and strong staining in the Hypertension and Hypertension + irisin groups (p<0.01). Control group (a); Irisin group (b); Hypertension group (c); Hypertension + irisin group (d).

Article Snippet: The immunohistochemical antibody [irisin (Bioss FNDC5 BS-4886-R), inducible NOS (Spring REF E3744), and endothelial NOS (Neomarkers RB-9279-P)] panel used for the diagnosis and differential diagnosis varies according to the cases and cytokeratin.

Techniques: Immunostaining, Staining

Journal: Balkan Medical Journal

Article Title: The Effects of Irisin on Nω-Nitro-L-arginine Methyl Ester Hydrochloride-Induced Hypertension in Rats

doi: 10.4274/balkanmedj.galenos.2019.2019.5.113

Figure Lengend Snippet: a-d. Glomerular immunostaining for neuronal nitric oxide synthase in different groups (x200). All groups viewed had similar widespread staining of glomerular capillaries (red arrow) and proximal tubules (blue arrow). Control group (a); Irisin group (b); Hypertension group (c); Hypertension + irisin group (d).

Article Snippet: The immunohistochemical antibody [irisin (Bioss FNDC5 BS-4886-R), inducible NOS (Spring REF E3744), and endothelial NOS (Neomarkers RB-9279-P)] panel used for the diagnosis and differential diagnosis varies according to the cases and cytokeratin.

Techniques: Immunostaining, Staining

Journal: Balkan Medical Journal

Article Title: The Effects of Irisin on Nω-Nitro-L-arginine Methyl Ester Hydrochloride-Induced Hypertension in Rats

doi: 10.4274/balkanmedj.galenos.2019.2019.5.113

Figure Lengend Snippet: a-d. Glomerular immunostaining for neuronal irisin in different groups (x200). All groups had staining with similar features in the glomerular capillaries (red arrow) and the distal tubules (blue arrow). Control group (a); Irisin group (b); Hypertension group (c); Hypertension + irisin group (d).

Article Snippet: The immunohistochemical antibody [irisin (Bioss FNDC5 BS-4886-R), inducible NOS (Spring REF E3744), and endothelial NOS (Neomarkers RB-9279-P)] panel used for the diagnosis and differential diagnosis varies according to the cases and cytokeratin.

Techniques: Immunostaining, Staining