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Figure 4


Fig. 4. Bcl-2 does not cooperate with CED-9 to inhibit CED-4-dependent yeast death. Yeast were transformed with the specified plasmids and spotted on inducing or repressing medium, as described in the legend to Fig. 1. FLAG-tagged CED-9 was expressed either by the constitutive ADH promoter or an inducible copper promoter (CUP1). The copper concentration in the inducing medium was varied to control FLAG-CED-9 expression levels in yeast transformed with the pCUP1-(TRP1)-FLAG-CED-9 plasmid. In the absence of added copper, only weak protection was afforded. The impact of Bcl-2 on this weak protection was tested by co-expressing Bcl-2, either using the intermediate-strength promoter (ADH) or the very strong promoter (GALL). Transformants were also grown in liquid galactose-containing media with no added copper. Lysates generated from those cultures were immunoblotted to visualise FLAG-CED-9 and Bcl-2 levels. Coomassie staining allowed visualisation of protein loading.





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