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. 2019 Oct 17;8:e41244. doi: 10.7554/eLife.41244

Figure 3. Piwi4 binds specifically to antisense v-piRNAs.

(A) Enrichment of SINV-derived small RNA in Piwi4-immunoprecipitation (IP) fraction compared to input sample. (B) The base bias for each position of SINV-derived piRNAs co-immunoprecipitated with Piwi4-FLAG shows a Uracil bias at position 1, characteristic of antisense Piwi-associated piRNAs (shown in bits). (C) Same as in A but for transposons (TE)-derived small RNAs.

Figure 3—source data 1. Identification of small RNAs preferentially bound by Piwi4 following Sindbis virus infection in the Aedes aegypti cell line Aag2.
Aag2 cells were transfected with plasmids expressing eGFP or FLAG-tagged Piwi4. Twenty-four hours after transfection, cells were infected with Sindbis virus and collected 3 days after infection and processed for IP of FLAG-Piwi4.
elife-41244-fig3-data1.xlsx (64.7KB, xlsx)
DOI: 10.7554/eLife.41244.010

Figure 3.

Figure 3—figure supplement 1. Piwi4 binds to bona-fide v-piRNAs and interacts with known siRNA and piRNA pathway components.

Figure 3—figure supplement 1.

(A) Results from two independent biological replicates (Exp1 and Exp2, left and right panels, respectively) for the enrichment of SINV-derived small RNA in Piwi4-IP fraction compared to input sample. (B) Lack of enrichment of SINV-derived small RNA in control eGFP immunoprecipitation (IP) fraction compared to input sample. (C) Network of Piwi4 protein interactions identified by affinity purification followed by mass-spectrometry using a C-terminal FLAG tagged-Piwi4 expressed in Aag2 cells as bait. Proteins known to be involved in piRNA pathways or in antiviral response in insects are shown in green or blue, respectively.