Journal: Scientific Reports

Article Title: In vitro exercise model using contractile human and mouse hybrid myotubes

doi: 10.1038/s41598-019-48316-9

Figure Lengend Snippet: Contraction-inducible upregulation of the expressions and secretions of myokines. After 7–8 days of differentiation, the differentiated myotubes originating from human samples alone (HSMM), human-origin and mouse-origin cells in a mixture (HSMM + C2C12), and murine C2C12 cells alone were treated with or without EPS at 20 V/25 mm, 1 Hz, 4-ms duration for 16 h. ( A ) Total RNA was extracted and the relative abundances of mRNAs for mouse IL-6 and CXCL1 as well as human IL-6 and CXCL1 were evaluated by real-time PCR analysis. Data normalized using mouse GAPDH or human RPLP0 transcripts were averaged over 3 independent experiments ( *P < 0.05). ( B ) Conditioned media were collected and concentrations of human IL-6 and CXCL1 as well as mouse IL-6 and CXCL1 were evaluated employing the BioPlex assay ( *P < 0.05, n = 3).

Article Snippet: The mouse CXCL1 and IL-6 concentrations were measured employing a house-made BioPlex assay with antibodies from the commercially available ELISA kits (R & D Systems, Minneapolis, MN).

Techniques: Real-time Polymerase Chain Reaction, BioPlex Assay