Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: Circulating exosomal miR-214-3p regulated the function of EPC in both humans and rats with obesity. (A and B) Targeted transcriptome sequencing of miRNAs within circulating exosomes derived from (A) humans and (B) rats unveiled the regulatory impact of aerobic exercise on miRNA expression profiles in these vesicles. The left panel shows a volcano plot of differentially expressed genes, and the right panel shows a heatmap representation of the transcriptome sequencing data, illustrating the expression patterns of differentially expressed genes in circulating exosomes. (C and D) qPCR was used to validate the relative expression levels of miR-214-3p in circulating exosomes derived from (C) humans ( n = 3 for each group; * p < 0.05, Exercise vs . Control) and (D) rats ( n : 6–12 for each group; * p < 0.05, HC vs . NC; # p < 0.05, HE vs . HC). (E) qPCR validation of miR-214-3p expression in miR-214-3p-overexpressing EPC ( n = 3 for each group). ** p < 0.01, miR-214-3p mimics vs . mimics NC; ## p < 0.01, miR-214-3p mimics vs . Control. (F) Summary data for cell vitality level among groups ( n : 10–12 for each group). *** p < 0.001, miR-214-3p mimics vs . mimics NC. (G and H) Summary data (G) and representative images (H) of wound healing in the scratch assay, showcasing the migration level of EPC among groups ( n = 4 for each group). * p < 0.05, miR-214-3p mimics vs . mimics NC. EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; mimics NC = mimics negative control; miR = microRNA; NC = the normal diet with sedentary group; qPCR = quantitative polymerase chain reaction.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: Sequencing, Derivative Assay, Expressing, Control, Biomarker Discovery, Wound Healing Assay, Migration, Negative Control, Real-time Polymerase Chain Reaction

Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: miR-214-3p plays an essential role in exercise-mediated protection against obesity-induced EPC dysfunction in vivo . (A) Dynamic weight change curves of rats. After randomization at Week 10, rats underwent treadmill exercise until the end of Week 18 ( n : 5–7 for each group). * p < 0.05, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC; §§ p < 0.01, KO + HE vs . WT + HE. (B) Summary data for Lee's index among groups at Week 10 and Week 18 ( n : 5–7 for each group). Left panel: ** p < 0.01, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; Right panel: *** p < 0.001, KO + HE vs . WT + HC; ### p < 0.001, WT + HC vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC. (C) Summary data for acetylcholine (ACh)-induced, endothelium-dependent relaxation in mesenteric arteries among groups ( n : 5–6 for each group). * p < 0.05, WT + HE vs . KO + HE; # p < 0.05, ## p < 0.05, WT + HE vs . WT + HC. (D) Summary data for EC 50 values in mesenteric arteries among groups in response to ACh ( n : 5–6 for each group). * p < 0.05, KO + HE vs . WT + HE; ## p < 0.01, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (E) Cell proliferation assays demonstrated that exosomes derived from the WT + HC group exhibited a diminished capacity to promote EPC proliferation compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC proliferation, which was abolished by knocking out miR-214-3p in rats ( n = 10 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (F) Scratch assay results showed that exosomes derived from the WT + HC group exhibited a diminished capacity to enhance EPC migration rates compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC migration rates, which was abolished by knocking out miR-214-3p in rats ( n = 6 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (G) Representative images of wound healing in the scratch assay, showcasing the migratory response of rat EPC. (H) qPCR analyses of pre-miR-214-3p and miR-214-3p in tissues from obese rats with and without exercise training ( n : 4–6 for each group). *** p < 0.001, HE vs . HC. EC 50 = half maximal effective concentration; EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; KO + HE = the knockout + high-fat diet with exercise group; miR = microRNA; pre-miR = precursor microRNA; WT + HC = the wild-type + high-fat diet with sedentary group; WT + HE = the wild-type + high-fat diet with exercise group; WT + NC = the wild-type + normal diet with sedentary group.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: In Vivo, Derivative Assay, Wound Healing Assay, Migration, Concentration Assay, Knock-Out

Journal: Journal of Animal Science and Biotechnology

Article Title: ZBED6 deletion activates FNDC1 -mediated PI3K/AKT and MAPK/ERK1/2 signaling pathways to promote myogenesis in pigs

doi: 10.1186/s40104-026-01504-w

Figure Lengend Snippet: Primer and siRNA sequences used in this study

Article Snippet: qPCR was performed on a CFX96 Real-Time PCR Detection System (Bio-Rad) in 20 μL reactions containing 10 μL of ChamQ SYBR qPCR Master Mix (Vazyme, Q311-02), 1 μL of diluted cDNA, 0.6 μL of each primer, and 7.8 μL of nuclease-free water.

Techniques: Sequencing

Journal: Genes to Cells

Article Title: A YAP – TEAD – CTCF Transcriptional Regulatory Axis Associated With the Elimination of Injured Hepatocytes From the Liver

doi: 10.1111/gtc.70150

Figure Lengend Snippet: Transcriptome analyses of livers and hepatocytes following YAP induction by AAV or HTVi. (a) Schematic overview of the RNA‐seq workflow for whole liver RNA‐seq (left) and sorted hepatocyte RNA‐seq (right). For sorted RNA‐seq, hepatocytes were isolated by collagenase perfusion and hepatocyte fractionation, followed by purification of live hepatocytes and FACS sorting of tdTomato‐positive hepatocytes. Liver tissues or sorted hepatocytes were collected 3 days after YAP induction and subjected to RNA‐seq analysis. (b) Unsupervised hierarchical clustering heatmap of whole liver RNA‐seq data using differentially expressed genes (DEGs) across the indicated groups (No treatment, HTVi‐empty, HTVi‐YAP, and AAV‐YAP). Red and blue indicate relatively high and low expression, respectively. (c) Venn diagram showing the overlap of significantly upregulated genes (fold change > 2, p < 0.05) in the AAV‐YAP and HTVi‐YAP groups compared with the no‐treatment control group in whole liver RNA‐seq. (d) Unsupervised hierarchical clustering heatmap of sorted hepatocyte RNA‐seq data using DEGs across the indicated groups (No treatment, HTVi‐empty, HTVi‐YAP, and AAV‐YAP). Red and blue indicate relatively high and low expression, respectively. (e) Venn diagram showing the overlap of significantly upregulated genes (fold change > 2, p < 0.05) in the AAV‐YAP and HTVi‐YAP groups compared with the no‐treatment control group in sorted hepatocyte RNA‐seq. (f) Venn diagram showing the overlap of HTVi‐specifically upregulated genes between whole RNA‐seq analysis and sort RNA‐seq analysis. (g) Reactome pathway analysis of genes commonly upregulated in the HTVi‐YAP group in both whole liver and sorted hepatocyte RNA‐seq datasets. The x ‐axis indicates −log10 (adjusted p value), dot size represents gene count, and color intensity indicates z‐score. (h–j) qPCR validation of representative genes associated with the indicated Reactome pathway terms: Cell cycle checkpoints (h), RHO GTPase effectors (i), and Immunoregulatory interactions between a lymphoid and a nonlymphoid cell (j). Relative mRNA expression was normalized to Gapdh . Data are shown as mean ± SD ( n = 3).

Article Snippet: cDNA was synthesized using ReverTra Ace qPCR RT Master Mix (Toyobo, Osaka, Japan).

Techniques: RNA Sequencing, Isolation, Fractionation, Purification, Analysis, Expressing, Control, Biomarker Discovery

Journal: Genes to Cells

Article Title: A YAP – TEAD – CTCF Transcriptional Regulatory Axis Associated With the Elimination of Injured Hepatocytes From the Liver

doi: 10.1111/gtc.70150

Figure Lengend Snippet: Transcriptome analyses of livers and hepatocytes following YAP induction by AAV or HTVi. (a) Schematic overview of the RNA‐seq workflow for whole liver RNA‐seq (left) and sorted hepatocyte RNA‐seq (right). For sorted RNA‐seq, hepatocytes were isolated by collagenase perfusion and hepatocyte fractionation, followed by purification of live hepatocytes and FACS sorting of tdTomato‐positive hepatocytes. Liver tissues or sorted hepatocytes were collected 3 days after YAP induction and subjected to RNA‐seq analysis. (b) Unsupervised hierarchical clustering heatmap of whole liver RNA‐seq data using differentially expressed genes (DEGs) across the indicated groups (No treatment, HTVi‐empty, HTVi‐YAP, and AAV‐YAP). Red and blue indicate relatively high and low expression, respectively. (c) Venn diagram showing the overlap of significantly upregulated genes (fold change > 2, p < 0.05) in the AAV‐YAP and HTVi‐YAP groups compared with the no‐treatment control group in whole liver RNA‐seq. (d) Unsupervised hierarchical clustering heatmap of sorted hepatocyte RNA‐seq data using DEGs across the indicated groups (No treatment, HTVi‐empty, HTVi‐YAP, and AAV‐YAP). Red and blue indicate relatively high and low expression, respectively. (e) Venn diagram showing the overlap of significantly upregulated genes (fold change > 2, p < 0.05) in the AAV‐YAP and HTVi‐YAP groups compared with the no‐treatment control group in sorted hepatocyte RNA‐seq. (f) Venn diagram showing the overlap of HTVi‐specifically upregulated genes between whole RNA‐seq analysis and sort RNA‐seq analysis. (g) Reactome pathway analysis of genes commonly upregulated in the HTVi‐YAP group in both whole liver and sorted hepatocyte RNA‐seq datasets. The x ‐axis indicates −log10 (adjusted p value), dot size represents gene count, and color intensity indicates z‐score. (h–j) qPCR validation of representative genes associated with the indicated Reactome pathway terms: Cell cycle checkpoints (h), RHO GTPase effectors (i), and Immunoregulatory interactions between a lymphoid and a nonlymphoid cell (j). Relative mRNA expression was normalized to Gapdh . Data are shown as mean ± SD ( n = 3).

Article Snippet: Quantitative real‐time PCR was performed using THUNDERBIRD SYBR qPCR Mix (Toyobo) on a CFX96 Real‐Time PCR System (Bio‐Rad, Hercules, California, USA).

Techniques: RNA Sequencing, Isolation, Fractionation, Purification, Analysis, Expressing, Control, Biomarker Discovery

Journal: Advanced Science

Article Title: ELF5‐Mediated Enhancer–Promoter Interaction Regulates LALBA Expression in Ovine Mammary Gland

doi: 10.1002/advs.78138

Figure Lengend Snippet: The relationship between chromatin accessibility and gene expression obtained from RNA‐seq. GO analysis of the up‐regulated (A) and down‐regulated genes (B) identified in ovine mammary gland between the two groups. KEGG analysis of the up‐regulated (C) and down‐regulated genes (D) identified in ovine mammary gland. Up‐regulated and down‐regulated means higher and lower expression in the high (H) lactation performance ewes compared to low (L) lactation group, respectively. (E) RT‐qPCR validation of DEGs in mammary gland using RNA‐seq. (F) Correlation analysis results between chromatin openness and gene expression. Each dot represents accessible chromatin regions (ACRs)‐gene pair, with the x‐axis showing the log 2 Fold Change in gene expression and the y‐axis showing the corresponding log 2 Fold Change in chromatin accessibility. (G) Visualization of DEGs driven by ACRs in mammary gland of H and L groups. The peaks are ACRs shown to provide genomic context, whereas the boxed regions were selected as representative differentially ACRs identified by ATAC‐seq, together with their corresponding regions in the RNA‐seq tracks.

Article Snippet: ChIP‐qPCR was performed to validate the occupancy of ELF5 and histone marks at the target genomic regions using SimpleChIP Universal qPCR Master Mix (CST, MA, USA; Cat. No.88989).

Techniques: Gene Expression, RNA Sequencing, Analysis, Expressing, Quantitative RT-PCR, Biomarker Discovery

Journal: Advanced Science

Article Title: ELF5‐Mediated Enhancer–Promoter Interaction Regulates LALBA Expression in Ovine Mammary Gland

doi: 10.1002/advs.78138

Figure Lengend Snippet: Identification of transcription factors (TFs) and analysis of ELF5 genomic binding in mammary gland of high (H) and low (L) lactation performance ewes. (A) Top 10 known motifs of the differential peaks between the two groups. (B) The top 10 TFs in terms of expression levels in the E26 transformation‐specific (ETS) family ( n = 3). (C) The expression of three epithelium‐specific ETS (ESEs) family members detected by RT‐qPCR ( n = 3). (D) Accumulated footprint plots for TFs of three ESEs family members in ovine mammary gland. (E) Functional annotation analysis results of the target genes closest to ELF5 peaks was shown by Venn diagrams. (F) Distribution of ELF5 peaks in different genomic functional regions. Data are represented as mean ± SD. Statistical significance was determined by two‐tailed unpaired t ‐test. ** p < 0.01.

Article Snippet: ChIP‐qPCR was performed to validate the occupancy of ELF5 and histone marks at the target genomic regions using SimpleChIP Universal qPCR Master Mix (CST, MA, USA; Cat. No.88989).

Techniques: Analysis, Binding Assay, Expressing, Transformation Assay, Quantitative RT-PCR, Functional Assay, Two Tailed Test

Journal: Advanced Science

Article Title: ELF5‐Mediated Enhancer–Promoter Interaction Regulates LALBA Expression in Ovine Mammary Gland

doi: 10.1002/advs.78138

Figure Lengend Snippet: Characterization and activity verification of LALBA super‐enhancer. (A) Expression of LALBA in mammary gland of high (H) and low (L) lactation ewes at mRNA and protein level. Representative images from three independent biological replicates are shown (Original uncropped blots can be found in Supplementary figure: “Original blots for (A)”). (B) Visualization of active histone marks at the LALBA super‐enhancer. (C) H3K27ac enrichment at enhancers of LALBA assessed by ChIP‐qPCR. (D) The activity of enhancers validated via luciferase assay. The LALBA mRNA (E) and protein expression (F) was determined when E4 activity was repressed using dCas9‐KRAB‐sgRNA. The “dCas9‐KRAB only” does not include an sgRNA. Original uncropped blots can be found in Supplementary figure: “Original blots for (F)”. Data are represented as mean ± SD ( n = 3). For statistical analysis, two‐tailed unpaired Student's t ‐tests were performed in A and C; one‐way ANOVA plus Tukey's post hoc tests were performed in (D–F). * p < 0.05, ** p < 0.01.

Article Snippet: ChIP‐qPCR was performed to validate the occupancy of ELF5 and histone marks at the target genomic regions using SimpleChIP Universal qPCR Master Mix (CST, MA, USA; Cat. No.88989).

Techniques: Activity Assay, Expressing, ChIP-qPCR, Luciferase, Analysis, Two Tailed Test