SPAG6 interacts with MYO1D and promotes the translocation of MYO1D to the plasma membrane. (A) A schematic representation of mass spectrometry detection. (B) Immunoprecipitation (IP) assay was carried out using SPAG6 antibody or IgG (negative control antibody). Samples were electrophoresed and Coomassie Brilliant Blue stained. Arrows indicate SPAG6 and the target bands. (C) Venn diagram of the protein strip and the protein lysate detected by LC-MS/MS analysis. (D) Interaction between SPAG6 and MYO1D was demonstrated via Co-IP in K562, HEL, HL-60, and THP-1 cells. (E) Representative images showing immunofluorescence staining of SPAG6 (red), MYO1D (green), and DAPI (blue) in K562 and HEL cells with treatment as indicated. Scale bar, 25 μm. (F) Western blot verified the different distribution of MYO1D in leukemia cells. (G) Western blot demonstrated SPAG6 expression with knockdown or overexpression of MYO1D in 4 AML cells. Co-IP, co-immunoprecipitation.