Cellular Lipid Extraction and Cholesterol Analysis
Cellular lipids were extracted using the method of Folch et al. [22 (link)] and cholesteryl esters were quantified using an enzymatic Amplex Red® Cholesterol kit (Thermo Fisher Scientific) [23 (link)]. Cell protein content was determined using Pierce Micro BCA protein assay (Life Technologies Australia Pty Ltd., Scoresby VIC, Australia). Media cholesterol was measured using the Amplex Red® Cholesterol kit according to manufacturer’s instructions (Thermo Fisher Scientific) [23 (link)]. Media testosterone was determined by the Department of Chemical Pathology, Royal Prince Alfred Hospital (Sydney, NSW, Australia). Neutral cholesterol ester hydrolase activity was determined as previously described [24 (link)].
Other organizations :
Heidelberg University, Universidade Federal do Espírito Santo, University of Adelaide, South Australian Health and Medical Research Institute, Freemasons Foundation Centre for Men's Health, UNSW Sydney, Garvan Institute of Medical Research, The Kinghorn Cancer Centre, St Vincent's Hospital
Method of Folch et al. for cellular lipid extraction
dependent variables
Cholesteryl esters quantification using Amplex Red® Cholesterol kit
Cell protein content determination using Pierce Micro BCA protein assay
Media cholesterol measurement using Amplex Red® Cholesterol kit
Media testosterone determination by the Department of Chemical Pathology, Royal Prince Alfred Hospital
control variables
Neutral cholesterol ester hydrolase activity determined as previously described
controls
Positive control: Not specified
Negative control: Not specified
Annotations
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