Journal: International Journal of Molecular Sciences

Article Title: Association of Irisin/FNDC5 with ERRα and PGC-1α Expression in NSCLC

doi: 10.3390/ijms232214204

Figure Lengend Snippet: Clinicopathological characteristics of patients with non-small cell lung cancer (NSCLC) related to low and high expression of irisin/FNDC5 (Chi 2 test analysis), significance in bold.

Article Snippet: The 7900HT Fast (Applied Biosystems, Waltham, MA, USA) Real-Time PCR System and the relative quantification (RQ) method were used to analyze the FNDC5 mRNA expression (FNDC5; Assay ID: Hs00401006_m1, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) and ESRRA mRNA expression (ESRRA; Assay ID: Hs00607062_gH, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) in cell lines and tissues.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Association of Irisin/FNDC5 with ERRα and PGC-1α Expression in NSCLC

doi: 10.3390/ijms232214204

Figure Lengend Snippet: Comparison between control (n = 16) and NSCLCs (n = 56) of FNDC5 mRNA ( A ) and ESRRA mRNA ( B ) expression levels. The moderate positive correlation between mRNA FNDC5 and mRNA ESRRA expression levels in NSCLC patients ( C ), * p ≤ 0.05, ** p ≤ 0.005.

Article Snippet: The 7900HT Fast (Applied Biosystems, Waltham, MA, USA) Real-Time PCR System and the relative quantification (RQ) method were used to analyze the FNDC5 mRNA expression (FNDC5; Assay ID: Hs00401006_m1, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) and ESRRA mRNA expression (ESRRA; Assay ID: Hs00607062_gH, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) in cell lines and tissues.

Techniques: Comparison, Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Association of Irisin/FNDC5 with ERRα and PGC-1α Expression in NSCLC

doi: 10.3390/ijms232214204

Figure Lengend Snippet: Comparison of the expression level of FNDC5 mRNA after co-culture in IMR-90 cells in the empty insert (control) ( A , B ) and the insert with lung cancer cells [NCI-H1703 ( C , D ) and NCI-H522 ( E , F )], * p ≤ 0.05, ** p ≤ 0.005, *** p < 0.001.

Article Snippet: The 7900HT Fast (Applied Biosystems, Waltham, MA, USA) Real-Time PCR System and the relative quantification (RQ) method were used to analyze the FNDC5 mRNA expression (FNDC5; Assay ID: Hs00401006_m1, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) and ESRRA mRNA expression (ESRRA; Assay ID: Hs00607062_gH, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) in cell lines and tissues.

Techniques: Comparison, Expressing, Co-Culture Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Association of Irisin/FNDC5 with ERRα and PGC-1α Expression in NSCLC

doi: 10.3390/ijms232214204

Figure Lengend Snippet: Positive immunogold reaction (black dots—indicated by arrows) point to irisin/FNDC5 expression in the cell cytoplasm in NCI-H522 cells—magnification on the right ( A ), in NCI-H1703 cell mitochondria-M membrane—magnification on the right ( B ), in rough endoplasmic reticulum-RER and in cytoplasmic extensions of A549 cell ( C ), N -nucleus, magnification ×25,000.

Article Snippet: The 7900HT Fast (Applied Biosystems, Waltham, MA, USA) Real-Time PCR System and the relative quantification (RQ) method were used to analyze the FNDC5 mRNA expression (FNDC5; Assay ID: Hs00401006_m1, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) and ESRRA mRNA expression (ESRRA; Assay ID: Hs00607062_gH, TaqMan Gene Expression Assay, Applied Biosystems, Waltham, MA, USA) in cell lines and tissues.

Techniques: Expressing, Membrane

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

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: Subsequently, the fixed cells were incubated with the specific polyclonal rabbit anti-irisin/FNDC5 antibody (dilution 1:50; code no. NBP2-14024; Novus Biologicals) at 4 °C overnight.

Techniques: Transmission Assay, Electron Microscopy, Double Staining

Journal: Bioengineered

Article Title: MicroRNA-665-3p exacerbates nonalcoholic fatty liver disease in mice

doi: 10.1080/21655979.2021.2017698

Figure Lengend Snippet: miR-665-3p antagomir activates AMPKα via increasing FNDC5 expression . (a) Graphic representation of the miR-665-3p binding motifs within the 3ʹ-UTR of FNDC5. (b-c) The mRNA and protein levels of FNDC5 in the liver from HFD mice. (d) The levels of hepatic irisin detected by a commercial ELISA kit. (e) Relative luciferase activity of the reporter constructs containing either WT or TRU 3ʹ-UTR of FNDC5 after treatment with miR-665-3p agomir. (f) The mRNA levels of FNDC5 in the liver from HFD mice treated with shFNDC5 or shRNA. (g) The levels of AMPKα phosphorylation. (h) Hepatic hydroxyproline levels. (i) Serum ALT and AST levels. All results were expressed as the means ± standard deviations, n = 6 for each group, and * P < 0.05 was considered statistically significant.

Article Snippet: Short hairpin RNAs against FNDC5 (shFNDC5) and scramble shRNA were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA), and then packaged into the liver-specific adeno-associated virus serotype 8 (AAV8) vectors by Shanghai GenePharma Co.,Ltd. (Shanghai, China).

Techniques: Expressing, Binding Assay, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Construct, shRNA, Phospho-proteomics

Journal: Bioengineered

Article Title: MicroRNA-665-3p exacerbates nonalcoholic fatty liver disease in mice

doi: 10.1080/21655979.2021.2017698

Figure Lengend Snippet: A diagram of the mechanisms of miR-665-3p in the pathogenesis of NAFLD . miR-665-3p directly binds to the 3ʹ-UTR of FNDC5 and inhibit its expression, thereby exacerbating oxidative stress and inflammation via inactivating AMPKα pathway during NAFLD.

Article Snippet: Short hairpin RNAs against FNDC5 (shFNDC5) and scramble shRNA were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA), and then packaged into the liver-specific adeno-associated virus serotype 8 (AAV8) vectors by Shanghai GenePharma Co.,Ltd. (Shanghai, China).

Techniques: Expressing

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – FNDC5 is depleted in the hearts of human cancer patients following chemotherapy exposure. (A) Representative Masson Trichrome staining and quantification in control (n = 10) and patients with a history of chemotherapy (n = 10) [scale bar = 100 μm]. (B) Representative immunostaining and quantification of cardiac FNDC5, ATR, and p73 in control (n = 10) and patients with a history of chemotherapy (n = 10) [scale bar = 100 μm]. (C) Representative immunoblots and quantification of cardiac FNDC5, γH2Ax, ATR, pChk1, p73, Bax, β-MHC, and ANP expression in controls and patients with a history of (n = 12/group). A detailed information regarding history of chemotherapy patients and controls is provided in β-Actin serves as a loading control for immunoblots. The data were analyzed by student's t-test or two-way ANOVA with Sidak's post-hoc test. ∗∗∗∗ P < 0.0001. The data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Staining, Control, Immunostaining, Western Blot, Expressing

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – FNDC5 overexpression in heart attenuates Doxorubicin-dependent ATR/Chk1 activation. (A) Representative immunoblot and quantification of FNDC5 expression in hearts isolated from Doxorubicin (9 mg/kg, i.p. biweekly; cumulative dose of 45 mg/kg) or saline treated mice (n = 8). (B) Irisin levels quantified in serum isolated from mice that received an intracardiac injection of FNDC5-targeted shRNA or a viral vector encoding FNDC5 (n = 12). (C) After the intracardiac injection of an FNDC5 encoding viral construct or vector control into mouse myocardium, mice (n = 6) received either Doxorubicin (9 mg/kg, i.p. biweekly; a cumulative dose of 45 mg/kg) or saline starting at 9 weeks of age. Samples for analyses were collected one week after the final Doxorubicin dose. Representative immunoblots and quantification of FNDC5, γH2Ax, ATR, pChk1, p73, ANP, and β-MHC expression. β-Actin serves as a loading control for immunoblots. Data were analyzed by student's t-test or one- or two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Over Expression, Activation Assay, Western Blot, Expressing, Isolation, Saline, Injection, shRNA, Plasmid Preparation, Construct, Control

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – The ability of FNDC5 to suppress ATR/Chk1 activation requires irisin signaling. (A–C) AC-16 cells were pre-treated with irisin (200 ng, 24 h), and scramble or FNDC5-specific shRNA was inserted. (A) Representative immunoblots and quantification of pChk1, p73, Bax, β-MHC, and ANP (n = 6). (B) Apoptosis (cytoplasmic histone-associated DNA fragments; n = 5). (D–F) AC-16 cells were transduced with plasmids encoding FNDC5 (FNDC5 overexpression, OE) or vector control, pre-treated with Cilengitide (10 μM, 24 h), and then exposed to Doxorubicin (3 μM, 16 h). (C) Representative immunoblots and quantification of pChk1, p73, Bax, βMHC, and ANP (n = 6). (D) Apoptosis (cytoplasmic histone-associated DNA fragments; n = 5). β-Actin serves as a loading control for immunoblots. Data were analyzed by one- or two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Activation Assay, shRNA, Western Blot, Transduction, Over Expression, Plasmid Preparation, Control

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – FNDC5 depletion increases oxidative stress and compromises mitochondrial function. AC-16 cells were transduced with either a plasmid encoding FNDC5 (FNDC5 overexpression, OE) or vector control or FNDC5-specific shRNA (FNDC5 KD) or scramble shRNA. (A) Baseline glutathione peroxidase (GPX) activity (n = 5) and superoxide dismutase (SOD) activity (n = 5). GPX and SOD activity (n = 5) in (B) FNDC5 KD or (C) FNDC5 OE cells treated with doxorubicin (3 μM, 16 h). (D) AC-16 cells expressing scramble or FNDC5-shRNA were pre-treated with irisin (200 ng, 24 h). GPX and SOD activity (n = 5) as well as CM-H 2 -DCFDA fluorescence (ROS; n = 5) are shown. (E) AC-16 cells were transduced with plasmids encoding FNDC5 (FNDC5 overexpression, OE) or vector control, pre-treated with Cilengitide (10 μM, 24 h), and then exposed to Doxorubicin (3 μM, 16 h). GPX and SOD activity (n = 5) as well as CM-H 2 -DCFDA fluorescence (ROS; n = 5) are shown. (F) AC-16 cells (n = 3) were transduced with a virus encoding FNDC5 (FNDC5 overexpression, OE) or vector control. Representative immunoblots and quantification of FNDC5, mitochondrial translated proteins (CV MT-ATP6, CIV MT-CO2, CIII MT-CYTB and CI MT-ND1) and nuclear translated proteins (CIII UQCRC2, CI NDUFB8 and CV ATP5A). (G–H) FNDC5 OE cells were treated with doxorubicin (3 μM, 16 h) and (G) mitochondrial Ca 2+ (n = 5) and (H) mitochondrial membrane potential (Δψ M , n = 5). β-Actin serves as a loading control for immunoblots. Data were analyzed by one- or two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Transduction, Plasmid Preparation, Over Expression, Control, shRNA, Activity Assay, Expressing, Fluorescence, Virus, Western Blot, Membrane

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – The ability of FNDC5 to decrease oxidative stress is not required for mitigation of Doxorubicin-dependent Chk1 activation. (A) AC-16 cells were transduced with plasmids encoding FNDC5 (FNDC5 overexpression, OE) or vector control, pre-treated with NAC (5 mM, 12 h), and then exposed to Doxorubicin (3 μM, 16 h). Representative immunoblots and quantification of pChk1(S296), pH2AX(S139), and FNDC5 are shown (n = 3). (D–E) AC-16 cells were transduced with vectors encoding scramble or lentiviral FNDC5-specific shRNA pre-treated with NAC (5 mM, 12 h). (B) Representative immunoblots and quantification of pChk1, p73, Bax, βMHC, and ANP (n = 6). (C) Apoptosis (cytoplasmic histone-associated DNA fragments; n = 5). β-Actin serves as a loading control for immunoblots. Data were analyzed by one- or two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Activation Assay, Transduction, Over Expression, Plasmid Preparation, Control, Western Blot, shRNA

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – FNDC5 forms a complex with Chk1 in VCM. (A) In silico modeling of the FNDC5-CHK1 supports a direct interaction. The best docking pose for the FNDC5-CHK1 complex was predicted using a ZDOCK molecular docking experiment. Some of the residues present on the protein-protein interaction interface are highlighted in red text labels. (B) Co-immunoprecipitation (Co-IP) of FNDC5 with Chk1 from human AC-16 cardiomyocytes (left panel). Co-IP of Chk1 with FNDC5 deletion constructs transfected into human AC-16 cardiomyocytes (right panel). Constructs lacking the irisin (Δ29-122), fibronectin III domain (Δ34-124), or hydrophobic transmembrane domain (Δ151-184) were tested for Chk1 binding. (C) Expression of FNDC5, Chk1, phosphorylated Chk1, Lamin A/C, and GAPDH in total cell lysates or the cyotosolic and nuclear fractions of AC-16 cells following either FNDC5 overexpression or knockdown (n = 3). Data were analyzed by one-way ANOVA with Sidak's post-hoc test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: In Silico, Immunoprecipitation, Co-Immunoprecipitation Assay, Construct, Transfection, Binding Assay, Expressing, Over Expression, Knockdown

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – FNDC5 depletion triggers cardiac dysfunction by increasing Chk1 activity. Scramble or lentiviral FNDC5-targeted shRNA were introduced into the murine myocardium via intracardiac injection. Administration of scramble or lentiviral FNDC5-shRNA via intracardiac injection. Beginning at age 9 weeks, the Chk1 inhibitor LY2606368 (250 mg/kg cumulative dose; 50 mg/kg biweekly for 10 weeks) or saline. Samples were collected 1 week after the last dose later for biochemical and histological analyses. (A) Representative immunostaining and quantification of cardiac FNDC5 and p73 (n = 10) [scale bar = 100 μm]. (B) Representative immunoblots and quantification of FNDC5, pChk1, p73, Bax, βMHC, and ANP (n = 6). (C) Masson Trichrome staining in heart tissue sections (n = 10) [scale bar = 100 μm]. (D) Left ventricular ejection fraction (LVEF) (n = 6). (E) Oxidative stress measures including GPX/SOD activity (n = 5) and CM-H 2 -DCFDA fluorescence (ROS; n = 5). (F) Survival curves of FNDC5-knockout and control wild-type mice. N = 18 in the wild-type group and N = 30 in the FNDC5-knockout group. β-Actin serves as a loading control for immunoblots. Data were analyzed by two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Activity Assay, shRNA, Injection, Saline, Immunostaining, Western Blot, Staining, Fluorescence, Knock-Out, Control

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – Cardiac FNDC5 is depleted with age resulting in increased susceptibility to cardiotoxic insult. (A) A lentiviral FNDC5 encoding viral construct or vector control was introduced into the mouse myocardium of one-month-old mice (n = 8). Beginning at age 2 months, mice received either Doxorubicin (9 mg/kg, i.p. biweekly; a cumulative dose of 45 mg/kg) or saline. Samples for analyses were collected one week after the final Doxorubicin dose. Representative immunoblots and quantification of FNDC5, γH2AX, pChk1, p73, ANP, and β-MHC are shown. (B) Representative immunoblotting analysis of FNDC5 expression in the hearts of young (2-month-old) and aged (24-month-old) mice (n = 8). (C) AC-16 & H9C2 cells were treated with Doxorubicin (0.1 μM, 24 h) for 5 days. FNDC5 protein expression in non-senescent (passage numbers 5–7) and replicative-senescent (passage numbers 25–30) were determined by immunoblotting analysis (n = 5). (D) Scramble or lentiviral FNDC5-targeted shRNA was introduced into the hearts of group of young (2 months) or aged (24 months) mice via intracardiac injection as described (n = 4). After shRNA introduction, hearts were collected for analysis. (E) Correlation between FNDC5 expression in heart and patient age in control (black) and chemotherapy-exposed patients (red). Representative immunoblots and quantification of γH2AX, ATR, and pChk1 are shown. β-Actin serves as a loading control for immunoblots. Data were analyzed by student's t-test, and Two-way ANOVA with Sidak's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns = not significant. Data are presented as mean ± SEM.

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Construct, Plasmid Preparation, Control, Saline, Western Blot, Expressing, shRNA, Injection

Journal: Redox Biology

Article Title: FNDC5/irisin mitigates the cardiotoxic impacts of cancer chemotherapeutics by modulating ROS-dependent and -independent mechanisms

doi: 10.1016/j.redox.2025.103527

Figure Lengend Snippet: – Schematic outlining the role of FNDC5 in Doxorubicin-induced cardiotoxicity. Our data support a model wherein FNDC5/irisin prevent Doxorubicin-dependent cardiomyocyte death by limiting ROS generation and activation of the mitochondrial apoptosis pathway and suppressing activation of the ATR/Chk1/p73 signaling axis. We have two hypotheses as to how FNDC5/irisin regulate Chk1: 1) Signaling via the irisin integrin receptor prevents Chk1 phosphorylation and activation or 2) FNDC5, possibly sequestered in the endocytic compartment, binds directly to Chk1 to block Chk1 phosphorylation. Image generated with biorender.com .

Article Snippet: To deplete FNDC5 in heart, 1 week old wild type mice received 5 × 10 8 lentiviral vector particles containing scramble or FNDC5-targeted small hairpin RNA (shRNA) (Santacruz Biotechnology, Paso Robles, CA, USA) in a 40 μl volume via intra-cardiac injection as previously described [ ].

Techniques: Activation Assay, Phospho-proteomics, Blocking Assay, Generated