Setup and implementation of a rapid mitochondria isolation technique for large-scale omic studies

Mechanical properties instruct cell behavior in physiological and pathological conditions, but our understanding of the underlying mechanisms remains to be elucidated. Recently, in our lab, we demonstrated that actomyosin contractility promotes phosphorylation of the fission factor MIEF1, limiting the recruitment of DRP1 at mitochondria, peri-mitochondrial F-actin formation, and mitochondrial fission. We found that mitochondrial fission is a general mechanotransduction mechanism that is sufficient and required to regulate three transcription factors of broad relevance such as YAP-TAZ, SREBP1/2, and NRF2 to control cell proliferation, lipogenesis, antioxidant metabolism, chemotherapy resistance, and adipocyte differentiation in response to mechanical cues. Here, we focus on and implement a rapid mitochondrial isolation technique based on immunoprecipitation to allow an omics quantitative analysis of the metabolic and lipid profile of mitochondria upon actomyosin tension changes. Our results would shed light on how mitochondria fulfill a unifying signaling function by which the mechanics of the tissue microenvironment coordinate complementary cell functions.