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. 2017 May 12;45(13):7736–7750. doi: 10.1093/nar/gkx379

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 6. — Target RNA binding is inhibited by SPMMV P1. 35S:TAS1c1, 35S:mGFP4, 35S:amiR173, 35S:HA-AGO1, 35S:HA-AGO1-DAH, 35S:Flag-P1_1–395 and 35S:P1-C88A/C103A respectively were infiltrated into N. benthamiana leaves as indicated. To detect protein–protein interactions native extracts were analyzed, while for RNA immunoprecipitations infiltrated leaves were crosslinked with formaldehyde prior to extract preparation. Input and IP protein fractions were analyzed by SDS-PAGE followed by anti-HA or anti-Flag Western blot to detect HA-AGO1/HA-AGO1-DAH or Flag-P1_1–395 proteins, respectively. Ponceau staining shows equal loading of proteins. Input and IP RNA fractions where analyzed by semi-quantitative RT-PCR to detect TAS1c or actin mRNAs. One representative blot from three biological replicates is shown.