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. 2019 Oct 16;4(5):e00580-19. doi: 10.1128/mSphere.00580-19

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Copyright © 2019 Zhang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 5 — EhAgo2-2 PIWI mutants have no effect on sRNA binding or protein localization. (A) sRNA binding ability is unaffected in three PIWI mutants. Overexpressing Myc-tagged cell lines of three PIWI mutants (EhAgo2-2 PIWI^D545A, EhAgo2-2 PIWI^N747A, and EhAgo2-2 PIWI^D545A;N747A) were used for anti-Myc IP pulldown experiments. The upper panel shows pCp-labeled IP sRNA for all cell lines, indicating that the bound sRNAs are unchanged for PIWI mutants. The lower panel shows anti-Myc Western blot analysis for the IP proteins. (B) EhAgo2-2 localization is unchanged in the overexpressing cell lines of three PIWI mutants. Anti-Myc IFAs for all three PIWI mutants show signal in the nucleus.