Additional file 1 of Effect of noradrenaline on propofol-induced mitochondrial dysfunction in human skeletal muscle cells
Creators: name: Krajčová, Adéla
name: Skagen, Christine
name: Džupa, Valér
name: Urban, Tomáš
name: Rustan, Arild C.
name: Jiroutková, Kateřina
name: Bakalář, Bohumil
name: Thoresen, G. Hege
name: Duška, František
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Physiology
subject: Evolutionary Biology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Table S1. Study subject characteristics on biopsy day. Figure S1. Cell viability. Results are expressed as the percentage of cell viability relative to the control (= non-treated cells). a) Individual groups represent viability of cells exposed for 96 h to different concentrations of noradrenaline. Data are presented a s the mean ± SEM (n = 4 subjects). Values for each experimental condition were measured in triplicates in each subject. b) Individual groups represent viability of cells exposed for 96 h to ethanol (= propofol vehicle; 0.1%), 0.1 mM noradrenaline alone and c) different concentrations (μg/mL) of either propofol alone or mixture of propofol and 0.1 mM noradrenaline. Data are presented as the mean ± SEM (n = 7 subjects). Values for each experimental condition were measured in triplicates in each subject. Note: NA = noradrenaline. *** p &lt; 0.001 vs. control group. Figure S2. Kinetic graph on XF24 Analyzer demonstrates changes after propofol at various concentrations. Real-time measurement of OCR at baseline and after sequential injection of oligomycin, FCCP and Antimycin A. Each data-point represents the mean of 7 independent samples (subjects) measured in tri- or tetraplicates normalized to protein content. Error bars indicate standard error of the mean. Different colours represent different groups exposed to propofol (0; 2.5; 10 µg/mL). Figure S3. A) Global mitochondrial parameters. Basal respiration, maximal respiratory capacity, ATP production and non-mitochondrial respiration. N = 7 replicates with 21–28 wells for each condition normalized to protein content. Error bars indicate standard error of the mean. B) Mitochondrial mass calculated as a fraction (%) of a cell surface area in 2D cross-sectional images. Figure S4. Uptake of CO2 production and lactic acid metabolism. A) Uptake of [14C]palmitic acid. B) [14C]lactic acid oxidation. C) [14C]lactic acid uptake. Note: NA = noradrenaline. Error bars in each graph indicate standard error of the mean. Note: NA = noradrenaline. Figure S5. Flow cytometry. Histogram showing MTG intensity of individual cell groups. Data are presented as the mean ± SEM (n = 2–3 experiments per each group). Note: NA = noradrenaline. Figure S6. Mitochondrial membrane potential. A) Myoblasts after staining with MitoTracker™ Green FM (left), TMRE (in the middle) and after staining of both agents (right). Experiments were performed at least at 60 cells per each group from n = 3 independent experiments (cells from 3 individual subjects). B) Determination of ∆ψm was expressed as TMRE/MTG ratio. The mitochondrial uncoupling agent FCCP was used as a positive control. Note: MTG = MitoTracker™ Green FM; TRME = tetramethylrhodamine ethyl ester; FCCP = carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone, NA = noradrenaline. Error bars indicate standard error of the mean. *p &lt; 0.05, ***p &lt; 0.001 vs. control group. Figure S7. Analysis of lipid droplets. A) Quantification of LD mass by cross-sectional area of BODIPY 493/503 normalized to cell area. B) LD size assessed by cross-sectional area of individual LDs. C) LD number normalized to cell area. Experiments were performed at least at 38 cells per each condition from 2 independent measurements (= 2 individual subjects). Error bars indicate standard error of the mean. ***p &lt; 0.001 vs. control group.