Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: Doxorubicin increases mitochondrial H 2 O 2 levels and causes cell death in rat cardiomyocytes. (A) H9c2 cells were treated with the indicated concentrations of DOX for 24 h. Cell death was assessed by Annexin V-FITC staining and analyzed using flow cytometer. Representative histograms of three independent experiments are shown. (B) H9c2 cells were treated with 1 μM DOX for the indicated times. Cell viability was measured using WST-1 reagent. (C-D) H9c2 cells were treated with 1 μM DOX for the indicated times and then stained with 5 μM CM-H 2 DCFDA (C) or 5 μM MitoPY-1 (D) for 15 min. Relative fluorescence intensity (RFI) was measured by flow cytometer. All data are presented as mean ± S.D. (n = 3). Statistical significance was determined by one-way ANOVA followed by Dunnett's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 compared to untreated control.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Staining, Flow Cytometry, Fluorescence, Control

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates mitochondrial H 2 O 2 accumulation and oxidative damage in DOX-treated cardiomyocytes. (A) H9c2 cells stably expressing either control (pSUPER) or PrxⅢ-targeting siRNA (pSUPER-siPrxⅢ) were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 9 h. Mitochondrial H 2 O 2 levels were assessed using the mitochondria-targeted fluorescent probe MitoPY-1 and visualized by green fluorescence. Scale bar, 75 μm. Five randomly selected microscopic fields per sample were analyzed and combined to represent one biological replicate. (B) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. FOXO3a localization was assessed by immunofluorescence using anti-FOXO3a (green) antibody and DAPI (blue). Scale bar, 25 μm Five randomly selected microscopic fields per sample were analyzed and combined to represent one biological replicate. All data are expressed as mean ± S.D. Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 and ns, p > 0.05.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Stable Transfection, Expressing, Control, Transduction, Fluorescence, Immunofluorescence

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates oxidative damage and mitochondrial bioenergetics in DOX-treated cardiomyocytes. (A) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Cells were labeled with 5 μM 10-NAO and relative fluorescence intensity (RFI) was analyzed by flow cytometer. Representative histograms and quantification are shown. (B) Cells were treated with 1 μM DOX for 12 h and labeled with 10 μM Rho-123 to assess mitochondrial membrane potential ( ΔΨ m ). The percentage of cells with low ΔΨ m was quantified by flow cytometry. Representative histograms and quantification are shown. (C) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Representative bar graph showing relative ATP concentration normalized to protein contents. (D-H) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. (D) Representative tracing of the oxygen consumption rate (OCR). Arrows indicate time points when cells were treated with oligomycin, FCCP, and rotenone plus antimycin A (Rot/AA), respectively. The OCR is calculated as (E) basal respiration, (F) maximal respiration, (G) spare respiration, and (H) proton-leak. All data are expressed as mean ± S.D. (n = 3-5). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Labeling, Fluorescence, Flow Cytometry, Membrane, Concentration Assay

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates mitochondrial biogenesis in DOX-treated cardiomyocytes. (A) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 6 h. Total RNA was isolated and the mRNA levels of PGC-1α, Nrf1, and mtTFA were determined by qRT-PCR. Representative bar graphs showing relative expression normalized to GAPDH are presented. (B) Relative mRNA expression of the mitochondrial DNA-encoded cytochrome c oxidase (COX) mRNA was measured as an indicator of mtDNA copy-dependent transcription. Representative bar graphs showing relative expression normalized to GAPDH are presented. (C) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. Representative immunoblots and quantitative analyses of PGC-1α, Nrf1, and mtTFA proteins. GAPDH and α-tubulin were used as loading controls. Densitometric quantification of protein expression was performed from three independent biological replicates. All data are presented as mean ± S.D. (n = 3). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Isolation, Quantitative RT-PCR, Expressing, Western Blot

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ protects cardiomyocytes against DOX-induced apoptotic cell death. Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 18 h. (A) Cell lysates were subjected to immunoblot analyses for cleaved Caspase-3 and cleaved PARP-1 with normalization to β-actin. Densitometric quantification of protein expression was performed from three independent biological replicates. (B) Apoptotic cell death was assessed by Annexin V-FITC and 7-AAD double staining followed by flow cytometric analysis. Quantification of Annexin V and/or 7-AAD-positive cells is shown. All data are presented as mean ± S.D. Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: Rat cardiomyocyte cell line, H9c2 cells (CRL-1446) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM high glucose (SH30022.01, Hyclone, Logan, UT, USA) supplemented with 10% FBS, 1% antibiotic-antimycotic, and 1 μg/ml puromycin.

Techniques: Transduction, Western Blot, Expressing, Double Staining

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 was downregulated in the AF mice. ( A ) Atrial tissue of mice was stained with hematoxylin and eosin (H&E) to display the morphological structure. CRIF1 expression in the atrial tissue of mice was detected using immunohistochemistry (200×). ( B ) The CRIF1 mRNA levels in the atrial tissue were detected by RT-qPCR. The mice cardiac muscle cell line HL-1 was treated with Ang II (0, 0.1, 0.2, 0.5, 1 and 2 μM) for 24 h. ( C ) The CRIF1 mRNA expression was determined by RT-qPCR. Data are presented as mean ± SD (n=8 mice in each group). t -test was applied. ***P<0.001 vs control group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Staining, Expressing, Immunohistochemistry, Quantitative RT-PCR, Control

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: Overexpression of CRIF1 activates the SIRT1/eNOS pathway in cardiomyocytes treated with Ang II. HL-1 cells were transfected with pcDNA3.1-CRIF1 (OE-CRIF1) or vector for 24 h, and then treated with Ang II (1 μM) for an additional 24 h. The mRNA expression levels of ( A ) CRIF1, ( B ) SIRT1 and ( C ) eNOS were evaluated using RT-qPCR. HL-1 cells were transfected with pcDNA3.1-CRIF1 (OE-CRIF1) or vector for 24 h, and were then cotreated with a SIRT1 inhibitor EX527 (10 μM) and Ang II (1 μM) for further 24 h. ( D ) The cell apoptosis was assessed by staining with TUNEL and DAPI, and observed under a fluorescence microscope (200 X). ( E ) Apoptosis was quantified by calculating TUNEL positive cells (normalized to DAPI-stained cells). Data are presented as mean ± SD in triplicates. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Staining, TUNEL Assay, Fluorescence, Microscopy, Control

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 overexpression inhibits Ang II–induced hypertrophy of HL-1 cells. ( A ) Cells were stained with α-actinin antibody and observed under a fluorescence microscope. Representative images are shown (200×). RT-qPCR was used to assess the mRNA expression levels of hypertrophy-related genes, including ( B ) ANP, ( C ) BNP, and ( D ) β-MHC. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Staining, Fluorescence, Microscopy, Quantitative RT-PCR, Expressing, Control, Plasmid Preparation

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 overexpression suppresses Ang II–induced inflammation in cardiomyocytes. ( A – C ) RT-qPCR was used to determine the mRNA levels of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6. ( D – F ) ELISA was used to measure the levels of TNF-α, IL-1β, and IL-6 in the culture medium of HL-1 cells. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: Overexpression of CRIF1 inhibits intracellular ROS generation and oxidative stress in cardiomyocytes treated with Ang II. ( A ) Cells were stained with DHE, and representative images of intracellular ROS are shown (200×). ( B ) The extent of intracellular ROS was quantified by counting DHE-positive cells (normalized to DAPI-stained cells). Cell lysates of HL-1 cells were used to detect the oxidative stress markers: ( C ) MDA, ( D ) SOD, ( E ) CAT, and ( F ) NO. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Staining, Control, Plasmid Preparation