Figure 5
Real time respirometry analysis on the effect of cholesterol treatment of pancreatic beta cells. (A) Oxygen consumption rate at 10 mM glucose in presence and absence of inhibitors that interferes mitochondrial respiration is measured in control as well as cholesterol accumulated BRIN-BD11 pancreatic beta cells with or without the pre-treatment of small molecule M1. (B) Cellular respiration rate in untreated and cholesterol treated pancreatic beta cells with or without the pre-treatment of M1. (C) Mitochondrial respiration derived through subtraction of non-mitochondrial respiration rate from cellular respiration rate in untreated and cholesterol treated pancreatic beta cells with or without the pre-treatment of M1. (D) Mitochondrial respiration dissected as proton leak and ATP linked respiration in untreated and cholesterol treated pancreatic beta cells with or without the pre-treatment of M1. (E) Maximal respiration as obtained after the addition of chemical uncoupler FCCP and after subtraction of non-mitochondrial respiration rates in untreated and cholesterol exposed pancreatic beta cells with or without the pre-treatment of M1. (F) Spare Respiratory Capacity obtained by subtraction of basal respiration rate from maximal respiration rates in untreated and cholesterol exposed pancreatic beta cells with or without the pre-treatment of M1. (G) Proton leak derived through subtraction of ATP linked respiration from mitochondrial respiration and expressed as percentage of total mitochondrial respiration, in untreated and cholesterol treated pancreatic beta cells with or without the exposure of small molecule M1. (H) Coupling efficiency, calculated as fraction of oxygen consumption sensitive to oligomycin, in untreated and cholesterol treated pancreatic beta cells with or without the exposure of M1. (I) Non mitochondrial respiration determined by using electron transport chain inhibitors, rotenone and antimycin in untreated and cholesterol treated pancreatic beta cells with or without the exposure of M1. (J) Extracellular acidification rate which gives an approximate estimation of glycolysis in untreated and cholesterol treated pancreatic beta cells with or without the pre-treatment of M1. All data are normalized for protein content , expressed as mean ± SEM of 3 independent experiments and presented as pmol O2/min (***P < 0.001 t-tests (unpaired)), ns: not significant. Chol: cholesterol.
