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Fig. 6. (A) The Mad1-Mad2 complex in the wild-type strain. A coimmunoprecipitation experiment using an anti-Mad2 antibody was performed in order to visualize the Mad1-Mad2 protein interaction (right). The total amount of proteins was analyzed by straight western blotting (left). Mad1 and Mad2 precipitates were analyzed using an anti-Mad2 antibody (lower panels), whereas Mad1-HA was blotted using an anti-HA antibody (top panels). Mad2 knockout strain ({Delta}mad2) was used as a negative control. (B) Localization of Mad2-GFP in the {Delta}mad1 strain. Mad2-GFP (middle) was observed in the wild-type strain (upper panel) or {Delta}mad1 strain (lower panel). DNA was stained with DAPI in the same cells (left). (C) Mad2-GFP expresses at the same level in both the wild-type and {Delta}mad1 strain. Mad2-GFP protein level was analyzed by western blot using an anti-GFP antibody (upper panel). GTPase was blotted using anti-Spi1 antibody as a loading control (lower panel). A non-tagged Mad2 strain was used as a negative control for Mad2.