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0], and 5?mM ethleneglycoltetraacetic-acid) for 1?h at 37��C. Luciferase assay The myostatin luciferase reporter gene recombinant plasmid was provided by Dr Allen.18 The Tranfectin Lipid Reagent (Bio-Rad, Hercules, CA) was used for the recombinant plasmid transfection to Venetoclax C2C12 myoblasts. When the cells are 80% confluent, the complexes of DNA (recombinant plasmid and Renilla internal reference, phRLTK- luc, Promega, Madison, WI) and transfection solutions were added to cells. After a 6?h incubation, myoblasts were treated with vehicle (PBS), ghrelin (1?��M), cisplatin (50?��M), and cisplatin (50?��M)?+?ghrelin (1?��M). For transient expression, assay for reporter gene activity was measured 24?h after transfection. The dual-luciferase reporter assay system (Promega) was used for detecting luciferase activity. Statistical analysis SPSS 18.00 Histone demethylase software for Windows (SPSS Inc., Chicago, IL) was used for all statistical analysis. Parameters are expressed as mean?��?standard error of the mean. Statistical comparisons were performed using one-way ANOVA followed by Tukey test adjustments for multiple comparisons. P values of 0.05 or smaller were considered statistically significant. The authors certify that they comply with the ethical guidelines for authorship and publishing of the Journal of Cachexia, Sarcopenia and Muscle.19 Results Effect of ghrelin on body weight, muscle mass, and myofiber size changes caused by LLC tumour or cisplatin Both LLC and cisplatin induced a significant decrease in body weight compared Enzalutamide mouse with control animals, whereas ghrelin administration prevented these changes (Figure ?AA and ?andB).B). Lean body mass was also decreased by LLC, and cisplatin and ghrelin also prevented these changes (for the LLC model, change from baseline for heat-killed?+?vehicle [HK?+?V] 95.70?��?4.30%, tumour?+?vehicle [T?+?V] 83.80?��?1.52%, and tumour?+?ghrelin [T?+?G] 96.47?��?2.04%, P?
