pDsRed2-Bid encodes a fusion of Discosoma sp. red fluorescent protein (DsRed2) and Bid, a member of the Bcl-2 family. In healthy, non-apoptotic cells, Bid resides in the cytosol as soluble protein. Upon induction of apoptosis, Bid translocates to mitochondria. In cells expressing the Bid-DsRed2 fusion, the translocation can be detected by fluorescence microscopy.
Monitoring Bid activation with the pDsRed2-Bid Vector. 3T3 cells were transiently transfected with pDsRed2-Bid (Panels A & B). Approximately 24 hr later, cells were incubated with 700 nM staurosporine, an apoptosis inducing agent, at 37°C for 3 hr. Cells were then fixed and examined with a Zeiss Axioskop equipped with the appropriate light filters. In untreated cells (Panel A), the Bid reporter (Bid-DsRed2) distributes uniformly in the cytosol, as expected. Following induction (Panel B), the reporter translocates to the surface of mitochondria, forming intensely fluorescent clusters—clear evidence that the Bid pathway has been activated.
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