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. 2019 Feb 6;10(2):172–187. doi: 10.1080/19490976.2018.1506668

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© 2019 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

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Figure 6. — Activated individual SifA domains restore SifA domain localization in caspase-3 KO Cells. Caspase-3 KO HeLa cells containing either V5-SifA-HA, V5-SifA or SifA-HA were infected with WT-SL1344. The cells were fixed and stained (refer to methods) at 1 and 8 hr post-infection. Without caspase-3, both domains of V5-SifA-HA remain in the perinuclear region at all time points following infection (top and bottom left panels), suggesting caspase-3 cleavage is important for domain localization (compare to top panels in Figure 5). When using V5-SifA, which resembles the post-caspase-3 cleavage form of the SifA N-terminal domain, we restore domain localization (top and bottom middle panels). Experiments were performed at least three times using cells of different passage. (b) Relative distance (measured using a micron scale) of the distance travelled by the N-terminal and C-terminal in both V5-SifA and SifA-HA at 1 and 8 hrs. post-infection. P Values (statistics calculated using unpaired Student’s t test): NS, not significant; **** P < 0.0001.