Figure 1.

MTUS1/ATIP1 was localized at the outer mitochondrial membrane in HNSCC cells. (A-D) HNSCC cells were subjected to immunofluorescent staining with MTUS1/Flag (green) and MitoTracker red staining (A) or COX IV (red) (B) or MFN1/2 (red) (C) or TOMM20 (red) (D) and counterstained with DAPI (blue). (E) Equal amounts of whole cell lysate, mitochondria and cytoplasmic, membrane, soluble nuclear and chromatin-bound extractions from HNSCC cells were analyzed by western blotting using the indicated antibodies including mitochondria marker (COX IV and Timm23), MOM marker (MFN2 and TOMM20) and Golgi marker (GM130). (F) Mitochondria were isolated and incubated with protease K for 5 min and then subjected to western blotting to detect ATIP1, mitochondria marker (COX IV and Timm23) and MOM marker (MFN2 and TOMM20). (G) Cell lysates from HNSCC cells transduced with lentivirus containing ATIP1-Flag were subjected to immunoprecipitation with an antibody against Flag, followed by western blotting with the indicated antibodies (MFN2 or TOMM20). (H) Cell lysates were subjected to western blotting with the MFN2 or TOMM20 antibodies in MTUS1/ATIP1-overexpressed or control HNSCC cells. Scale bar, 10 mm. β-actin or GAPDH was used as loading proteins.