CETSA was conducted using M-PER (R) Mammalian Protein Extraction Reagent (Solarbio, Beijing, China) for protein extraction. Cell extracts were incubated with either DMSO or Curculigoside/Chebulagic acid for 1 h at room temperature. Each sample was divided into 7 tubes and heated at a specific temperature for 5 min with gentle mixing. Following centrifugation at 12,000 g at 4 °C for 20 min, the resulting supernatants were analyzed using immunoblotting.
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