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Add Sample, Standard, Biotin-antibody, and Streptavidin-HRP by turn, with incubation and washing

In addition, FUNDC1 is acetylated and deacetylated at K104 by Tip60 and HDAC3, which regulates the dimerization of FUNDC1 and thus its ubiquitination by MARCH5 [116]

Traverso et al., 2013)

The samples were resolved on a 1015% polyacrylamide gel and scanned on a Typhoon variable mode imager or on a FluorChem M System (ProteinSimple, San Jose, CA, USA)

Western immunoblot analyses For the preparation of total liver homogenates, frozen liver pieces were homogenized in RIPA buffer (150 mM NaCl, 1% TritonX-100, 0.25% sodium deoxycholate, 0.1% SDS, 50 mM Tris, 1 mM EDTA, 1 mM PMSF, protease inhibitor cocktail, pH 7.4) on ice with a tissue tearor (BioSpec Products, Bartlesville)