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Fig. 2. Overview of HeLa cells transfected with pEGFP-TPPP/p25. (A) Cells were fixed in cold-buffered ethanol for immunocytochemistry with anti-tubulin antibody. The representative image shows three TPPP/p25-transfected cells with distinct morphologies in the same microscopic field. Note the colocalization (orange) of EGFP-TPPP/p25 (green) with the microtubular network (red) at low expression levels (cell marked 1). The other two cells with bright fluorescence show an aberrant microtubular network and morphological changes with colocalization (marked 2 and 3). Comparing the fluorescence of these three transfected cells suggests low (cell 1) and high (cells 2 and 3) levels of expressed protein. (B) TMRE uptake in transfected HeLa cells. TMRE (red) was used to investigate mitochondrial membrane state in live cells. At low levels of expression, the cells show a polarization state similar to that of untransfected cells. (C) HeLa cells transfected with pEGFP-C1 plasmid. Note the strong nuclear presence of the EGFP protein. (D-F) HeLa cells transfected with EGFP-TPPP/p25 plasmid. Representative examples of the different expression `classes' are presented: (D) low-level expressing; (E) aggresome-like; (F) nuclear cage. Bars, 10 µm.

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