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Fig. 2. WT.TACE-eGFP and T735A.TACE-eGFP proteins reside in the ER. (A) HeLa cells were transfected with 0.3 µg pWT.TACE-eGFP and pT735A.TACE-eGFP, fixed using methanol and stained for the ER-specific marker, calregulin (C-17) and Alexa 546 donkey anti-goat secondary antibodies. The coverslips were mounted and visualised using laser-scanning confocal microscopy. The TACE-eGFP protein, ER and nuclei are green, red and blue, respectively. (B) HeLa cells stably expressing WT.TACE (WT.HeLa) were seeded on coverslips and the following day fixed using methanol, stained for TACE using anti-TACE antibody (C-15) and Texas Red-conjugated donkey anti-goat secondary antibodies and the ER stained using anti-calregulin (H-170) and Alexa 488 donkey anti-goat secondary antibodies. Cells were visualised using laser-scanning confocal microscopy with the ER in green, TACE in red, nuclei (stained using DAPI) in blue and colocalisation highlighted with arrowheads (top panels). Untransfected HeLa cells were treated identically and stained using anti-TACE (C-15) primary antibody followed by Alexa 546 donkey anti-goat secondary antibodies. Red and blue channels for TACE and nuclei are shown, respectively (bottom panels). Bar, 10 µm.

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