In this research, to discover the interactions of mpox-A5 with host proteins, HEK293T cells were utilized and transfected with pCAGGS-HA-A5L plasmid (or pCAGGS-HA plasmid), harvested at 24h, Heterologous expression of MPXV A5L in HEK293T cells was performed for Co-IP with anti-HA beads, and Immunoblotting was conducted to verify the protein expression. The Co-IP products were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS).