Replication Data for: Fatty Acid oxidation of "Replicative Senescence in Mesenchymal Stem Cells: An In Vitro Study on Mitochondrial Dynamics and Metabolic Alterations"

Fatty Acid Oxidation Assay (ab222944, Abcam) was used to monitor FAO, the primary metabolic pathway for degradation of fatty acids, in living cells. bMSCs seeded on 96-well black plate (Costar, Sigma-Aldrich) (6 × 103 cells/well) were rinsed twice with Fatty Acid-Free medium added with 0.5 mM L-carnitine and 2.5 mM D-Glucose. In the meantime, Fatty Acid Measurement Medium was prepared by adding Oleate (FAO-conjugate) to Fatty Acid-Free Medium and added to each assay well. Extracellular O2 Consumption Reagent (λex/λem = 380/650 nm) was then added to the wells, and each well was sealed with pre-warmed High Sensitivity Mineral Oil. Treatment with the mitochondrial membrane potential uncoupler carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, 0.625 µM) was used as positive control. The FAO inhibitor Etomoxir (40 μM) was utilized as a negative control Fluorescence was acquired every 1.30 min for 90 min at Varioskan LUX Multimode Microplate Reader pre-set to 37 °C. Data were normalized to the cell number. The results are expressed as a box plot graph.