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The GST-pulldown assay was performed as previously described [31] with slight modifications. In brief, purified GST-tagged RSL1D1 protein was co-incubated with Glutathione Sepharose 4B beads at 4 °C for 1 h. Then, the beads were incubated with purified His-tagged p53 protein at 4 °C for 1 h, followed by washing five times with 1% Triton X-100 in PBS. The beads were boiled in SDS-PAGE sample loading buffer for western blot analysis.
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