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Updated: Sep 13, 2025

Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
Published on: March 23, 2015
Protocol for assessing mitochondrial cholesterol transport and protein molten globule state in steroidogenic and
Himangshu S Bose1, Mahuya Bose2, Randy M Whittal3
1Laboratory of Biochemistry and Cell Biology, Department of Biomedical Sciences, Mercer University School of Medicine, Savannah, GA 31404, USA; Anderson Cancer Institute, Memorial University Medical Center, Savannah, GA 31404, USA.
Abstract:
Steroid hormones are essential for the survival of all mammals for carbohydrate metabolism, stress management, and sexual reproduction. Here, we present a protocol for assessing mitochondrial cholesterol transport and protein molten globule state via measurement of pregnenolone or progesterone synthesis in steroidogenic and nonsteroidogenic cellular systems. We describe steps for cell culture, transfection, and measurement of steroidogenic activity from nonsteroidogenic cells. We then detail procedures for metabolic conversion. For complete details on the use and execution of this protocol, please refer to Bose,1 Bose et al.,2 Pawlak et al.,3 and Prasad et al.4.
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