HeLa–Tet-On–FLAG-SPINDOC-GFP cells were established for inducible expression and affinity purification. FLAG-SPINDOC-GFP expression was first induced by doxycycline treatment. Subsequently, cells were either left unsynchronized or arrested at the G2/M phase by nocodazole treatment. Efficient expression and enrichment of FLAG-SPINDOC-GFP were confirmed by FLAG co-immunoprecipitation. Immunoprecipitates were then subjected to mass spectrometry analysis to identify SPINDOC-associated proteins, particularly under mitotic conditions.