Fig. 10.

The pull-down assay conducted on HT22 cell lysate to confirm the physical interaction between melittin and Keap1. This assay confirmd the binding of melittin to Keap1 by capturing Keap1 protein complexes using melittin as bait. The presence of Keap1 in the pulled-down samples, extracted from melittin conjugated beads indicated a direct physical association between Keap1 and melittin. Tandem mass spectroscopy also confirmed the main attached protein was Keap1. This experimental validation corroborated the docking simulations and biochemical findings, providing robust evidence for the interaction between melittin and Keap1 at the protein level