Fig 4.

Stimulated interaction of the M-WT protein with the H protein after the disruption of F-actin. (A) Colocalization of the M proteins with the H protein after Cyt-D treatment. The M-WT or M-F50P protein was coexpressed with the H-myc protein. At 12 h posttransfection, the culture medium was replaced by medium including Cyt-D. At 24 or 48 h posttransfection, the cells were treated as described for Fig. 2B, and the distribution of the M and H proteins was observed under a confocal microscope. (B) Enhanced coprecipitation of the H protein with the M-WT protein after Cyt-D treatment. The M-WT or M-F50P protein was coexpressed with the H-myc protein in 293T cells. At 12 h posttransfection, culture medium was replaced by medium including Cyt-D or DMSO. At 24 h posttransfection, cell extracts were prepared by 2 ways with detergent solution and a Dounce homogenizer and subjected to immunoprecipitation as described in Fig. 2C. Proteins precipitated with anti-M antibody were analyzed by immunoblotting with anti-M protein mouse monoclonal antibody, anti-myc tag rabbit polyclonal antibody, anti-β-actin mouse monoclonal antibody, and anti-golgin97 mouse monoclonal antibody as the first antibodies.