2.7. Cellular thermal shift assay (CETSA)

THP‐1 cells were pretreated with or without 10 μM AEDC for 12 h. The cells were lysed by RIPA lysis buffer. The cell lysates were incubated in ice for 10 min and then centrifuged at 12,000 g for 10 min at 4°C. The protein concentration was determined using a BCA Protein Assay Kit and adjusted to 2 μg·μl⁻¹ using RIPA lysis buffer; 50‐μL cell lysates was transferred to new tubes and heated for 5 min at different temperature (50–90°C) using a thermal cycler. After incubation in ice for 10 min, soluble proteins were separated by centrifugation at 12,000 g for 20 min at 4°C and analysed by western blotting analysis (Jafari et al., 2014; Martinez et al., 2013).