Journal: eBioMedicine

Article Title: Exercise-derived peptide protects against pathological cardiac remodeling

doi: 10.1016/j.ebiom.2022.104164

Figure Lengend Snippet: The primers used in this study.

Article Snippet: The concentrations of CCDC80tide in EVs were measured by ELISA (Human CCDC80 ELISA Kit, EK1962, Boster Bio, USA) combined with immunoblotting assay.

Techniques:

Journal: eBioMedicine

Article Title: Exercise-derived peptide protects against pathological cardiac remodeling

doi: 10.1016/j.ebiom.2022.104164

Figure Lengend Snippet: CCDC80 is a potential exerkine involved in cardiovascular pathophysiology . a. Circos plot visualizes the overlap between upregulated gene lists from three databases, and the blue curves link genes that belong to the same enriched GO term. b. Pie charts represent the network of enriched terms in upregulated genes. Each circle node represents an enriched term and is clustered by its identity, where the size of a slice represents the percentage of genes under the term that originated from the corresponding database. c. Heatmap of enriched terms across the three upregulated gene lists, colored by p -values. d. Heatmap of enrichment analysis in DisGeNET, colored by p-values. e. Circos plot displays the overlap between upregulated gene lists, and the purple curves link identical genes. The five genes within the black box represent the genes with significant upregulation throughout the three databases. f. RT-PCR analysis of the expression levels of CCDC80 in various tissues of mice. g, h. After overexpression of PGC-1α in C2C12 cells, the expressions of Pgc-1α (g) and Ccdc80 (h) was determined by qPCR. i, j and k. Eight-week-old male mice were forced to swim 5 days per week for 3 months. Control groups mice were raised normally without swimming training. After complete swimming training, the skeletal muscles of mice were collected for western blot analysis of CCDC80 and PGC-1α ( n= 7). Data are presented as the average of four biological replicates (n = 4) ± SD. Statistical significance was determined using Student's t-test, *** p < 0.001.

Article Snippet: The concentrations of CCDC80tide in EVs were measured by ELISA (Human CCDC80 ELISA Kit, EK1962, Boster Bio, USA) combined with immunoblotting assay.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Over Expression, Control, Muscles, Western Blot

Journal: eBioMedicine

Article Title: Exercise-derived peptide protects against pathological cardiac remodeling

doi: 10.1016/j.ebiom.2022.104164

Figure Lengend Snippet: CCDC80 generates a peptide encapsulated in EVs in response to PGC-1α overexpression . a. The cell lysates and culture supernatants collected from vehicle- or Flag-PGC-1α-overexpressing C2C12 cells (treated or not with BFA, 1 μg/mL, 16 h) were prepared and analyzed for CCDC80 protein levels by WB. N= 3 b. The supernatants of CCDC80-EGFP-overexpressing C2C12 cells were harvested and filtered through a sterile 0.22-μm filter and then added to H9c2 cells. After 12-h incubation, the cell fluorescence was detected. The fluorescent spots indicated by white arrows were supposedly EVs. N= 3 c, d, e. The supernatants of CCDC80-overexpressing C2C12 cells were collected, and EVs were extracted for TEM (c), NTA (d), and EVs markers (e) determinations. N= 3. WCL: whole cell lysates. f. The cell lysates and EVs isolated from the supernatants of vehicle- or CCDC80-HA-overexpressing C2C12 cells were prepared and analyzed for CCDC80 protein levels by WB. g. The schematic diagram of CCDC80tide in CCDC80.

Article Snippet: The concentrations of CCDC80tide in EVs were measured by ELISA (Human CCDC80 ELISA Kit, EK1962, Boster Bio, USA) combined with immunoblotting assay.

Techniques: Over Expression, Sterility, Incubation, Fluorescence, Isolation