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Currently, this hypothesis is replaced by the multiple hit hypothesis that considers multiple factors (such as lipotoxicity, diet, sedentary lifestyle, epigenetics, insulin resistance, innate immune activation, environmental pollution, etc

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PhoA accumulation was analyzed by SDS-PAGE (NuPAGE 4 bis 12 %, Bis-Tris, Invitrogen, Waltham, MA, USA) and Western blot analyses were carried out using nitrocellulose membranes in the iBLOT2 system (Invitrogen, Waltham, MA, USA), both according to the supplier