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Molecular cloning of MAST4 and site-directed mutagenesis We used Phusion High-Fidelity DNA Polymerase to amplify the N-terminal (MAST4-N) and C-terminal (MAST4-C) halves of human MAST4 cDNA using the cDNAs of HEK293 cells as a template (primers are listed in Table S2)

To assess mitochondrial complex activities (specifically complex I, II and IV), the injection strategy was as follows: port A: rotenone (final concentration 2 M, Sigma-Aldrich), port B: succinate (final concentration 10 mM, cat.no

Palmitate preparation is a major source of variation in cell-based experiments

Baric et al., 2016

This approach maximizes stability compared to long-term mixture storage