2.2.9. Cellular thermal shift assay (CETSA)

Cells were collected and freeze-thawed three times using liquid nitrogen. Cell suspensions were divided into 2 aliquots and treated with or without J13 (5 μmol/L) for 1 h at room temperature. Each aliquot was heated at indicated temperature (38–64 °C) for 3 min followed by cooling for 3 min. Supernatants were isolated and analyzed by Western blot. Additionally, for living cell CETSA experiment, cells were treated with or without J13 (5 μmol/L) for 6 h, harvested, and heated separately at indicated temperature (38–64 °C) followed by 3 min cooling. Cells were collected and freeze-thawed three times using liquid nitrogen. Supernatants were analyzed by Western blot.