Whole cell protein lysates were prepared in RIPA Lysis buffer in the presence of protease and protein phosphatase inhibitors (Thermo Fisher) as previously described [17 (link)]. Briefly, whole cell lysates were vortexed every 10 min for 1 h on ice and centrifuged at 15,000 rpm for 5 min. The protein concentrations of the supernatants were determined by a BCA assay kit as per manufacturer’s instructions (Thermo Fisher).
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