Journal: Frontiers in Physiology

Article Title: Ion channels and atrial fibrillation: mitophagy as a key mediator

doi: 10.3389/fphys.2025.1687578

Figure Lengend Snippet: In vivo experimental validation. (A–F) The (A) electrocardiogram, (B) hematoxylin-eosin, (C) Masson, (D) quantification of collagen volume fraction, (E) Sirius staining, and (F) quantification of Sirius red staining area percentage in rats with acetylcholine-calcium chloride-induced atrial tachyarrhythmias ( n = 5). (G) Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to detect the relative expression levels of Hub genes, mitophagy, and ion channel-related genes ( n = 5). (H) Western blotting to detect the expression of BAX, TLR4, MIF, and GLUL proteins ( n = 3). Results are expressed as the mean ± S.D. * P < 0.05 vs. the control group, ** P < 0.01 vs. the control group. The normality of data distribution was assessed using the Shapiro-Wilk test, and the homogeneity of variances was verified by the Brown-Forsythe test. For comparisons among multiple groups that satisfied both normality and homogeneity of variances, a one-way analysis of variance (ANOVA) was employed, followed by Tukey’s honestly significant difference (HSD) post hoc test for specific group comparisons. For data that did not meet the assumptions of parametric tests, the non-parametric Kruskal-Wallis H test was used.

Article Snippet: Reagents: Acetylcholine Chloride (Ach) (Shanghai yuanye, Cat. S30170-5g); Anhydrous Calcium Chloride (CaCl 2 ) (Shanghai yuanye, Cat. S24110-500 g); Isoflurane (SAMVET, Lot.235180101); Hematoxylin-Eosin Constant Dye Kit (Servicebio, Cat. G1076); Masson Dye Solution Set (Servicebio, Cat. G1006); Differentiation Fluid (Servicebio, Cat. G1039); Sirius Red Staining Solution (Servicebio, Cat. G1018); Environmentally Friendly Dewaxing Liquid (Servicebio, Cat. G1128); Paraformaldehyde Fixative (Neutral) (Servicebio, Cat. G1101); 75% ethanol (Shenyang GuangCai, Cat.20250102); Absolute ethanol (Sinopharm Chemical Reagent Co.,Ltd., Cat.100092683); Xylene (Sinopharm Chemical Reagent Co.,Ltd., Cat.10023418); Normal Butanol (Sinopharm Chemical Reagent Co.,Ltd., Cat.100052190); Neutral Gum (Sinopharm Chemical Reagent Co.,Ltd., Cat.10004160); RNA Extraction Solution (Servicebio, Cat. G3013); chloroform Substitute (Servicebio, Cat. G3014); Isopropanol (Sinopharm Chemical Reagent Co.,Ltd., Cat.80109218); SweScript All-in-One RT SuperMix for qPCR (Servicebio, Cat. G3337); 2×Universal Blue SYnthetic Binding Reagent Green qPCR Master Mix (Servicebio, Cat. G3326); Bicinchoninic Acid Protein Concentration Measurement Kit (Beyotime, P0010); Radioimmunoprecipitation Assay Lysate (Beyotime, P0013B); Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) Gel Rapid Preparation Kits (Beyotime, P0015L); Maker (Epizyme Biotech, Cat. WJ103); β-Actin Rabbit mAb (Abclonal, Cat. AC038); Anti-MIF (UpingBio, Cat. YP-Ab-15949); Anti-GLUL (UpingBio, Cat. YP-Ab-12846); Anti-TLR4 (UpingBio, Cat. YP-Ab-13172); Anti-BAX (BOSTER, Cat. A00183); Goat Anti-Rabbit IgG (H + L), HRP (UpingBio, Cat. YP848537-H).

Techniques: In Vivo, Biomarker Discovery, Staining, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Control