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Cellular thermal shift assay (CETSA)
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The CETSA assay was performed as previously described [47]. Prostate cancer cells were treated with 50 μM ivermectin for 1 h. Cells were suspended in PBS with protease inhibitors, and heated at the indicated temperature for 3 min. Samples were subjected to three freeze-thaw cycles freeze-thaw using liquid nitrogen and centrifuged. Supernatants were collected and detected by western blot.
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