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Fig. 1. Overexpression of TAJ/TROY induces non-apoptotic cell death. (A) Colony formation after TAJ/TROY transfection. HEK293 and HeLa cells were transfected with an empty vector or expression vectors encoding Bax or TAJ/TROY. Overexpression of TAJ/TROY significantly decreased clonogenic survival of the cells. The figure shows a representative experiment with the means and standard deviations from triplicate plates. (B) Electron micrographs of TAJ/TROY-induced cell death. HEK293, HeLa and 293T cells were transfected with a vector control or plasmids encoding Bax or TAJ/TROY along with a GFP-encoding vector. Whereas Bax-transfected 293T cells (b) displayed classic characteristics of apoptosis, including chromatin condensation and apoptosis body, TAJ/TROY-transfected HEK293 (e), HeLa (g) and 293T (c) cells showed extensive cytoplasmic vacuolization and swelling of mitochondria or ER, along with absence of nuclear fragmentation, membrane blebbing or apoptotic-body formation. The following micrographs of empty-vector-transfected HEK293 (d), HeLa (f) and 293T (a) cells provided normal cell controls. Higher-magnification images of the vacuolization suggested that some vacuoles might be formed from the remnants of the mitochondrial (h) or ER (i). The transfection efficiency of the indicated plasmids ranged from 60% to 70% as determined by the expression of co-transfected GFP. Scale bars, 2 µM (a-g), 0.2 µM (h,i).

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