Whole-cell lysates were prepared in RIPA lysis buffer in the presence of protease and protein phosphatase inhibitors (Thermo Fisher) as previously described [15 (link)]. Lysates were vortexed every 10 min for 1 h at 4 °C and centrifuged at 15,000× g rpm. Protein concentrations of the supernatants were determined using the BCA assay kit as per the manufacturer’s instructions (Thermo Fisher).
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