Immunoprecipitation coupled with mass spectrometry (IP-MS) was performed to identify proteins interacting with ACTL8 in human cells. Cell lysates were immunoprecipitated using an anti-ACTL8 antibody or IgG control, followed by LC-MS/MS analysis on a Q Exactive HF Orbitrap mass spectrometer. Protein abundance was estimated based on peptide-spectrum matches (PSMs). Proteins enriched more than two-fold in ACTL8 immunoprecipitates compared with IgG controls and with p < 0.05 were considered high-confidence ACTL8-interacting proteins.