2.4 Cellular thermal shift assay (CETSA)
HEK-293T cells transfected with NF-κB plasmid and RAW 264.7 macrophages
were incubated with DMSO or 270 for 3 h. Cells were harvested and
resuspended in PBS and divided into four aliquots. Then cells were
heated at different temperatures by T100TM Thermal
Cycler (Bio-Rad). The heated cells were freeze-thawed three times with
liquid nitrogen and followed by centrifugation at 20,000×g for 20 min at
4 °C. The supernatants were harvested and loading buffer was added
before boiling. Protein levels were measured by western blotting.