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

Figure 2. Piwi4 is required for v-piRNA maturation.

(A) Size distribution plot of small RNAs mapping to the SINV genome from infected Aag2 cells treated with Fluc/control, Piwi4, or Ago3 dsRNA. (B) Size distribution plot of beta-elimination resistant small RNA mapping to the SINV genome from infected Aag2 cells treated with Fluc/control, Piwi4, or Ago3 dsRNA. (C) Same as in (A) but read counts were normalized to viral copy numbers. (D) Same as in (B) but read counts for beta-elimination resistant small RNAs were normalized to viral copy numbers.

Figure 2—source data 1. Identification of small RNAs in the Aedes aegypti cell line Aag2 knocked-down for Ago2, Ago2 or Piwi4 expression and infected with Sindbis virus.
Aag2 cells were transfected with dsRNA against Firefly luciferase (control), Ago2, Ago3 or Piwi4, infected with Sindbis virus 2 days later and collected 3 days after infection for small RNA extraction.
elife-41244-fig2-data1.xlsx (69KB, xlsx)
DOI: 10.7554/eLife.41244.007

Figure 2.

Figure 2—figure supplement 1. Effects of Ago2, Ago3 and Piwi4 knockdown on SINV-derived small RNAs.

Figure 2—figure supplement 1.

(A) Ago2, Ago3, and Piwi4 mRNA measured by RT-qPCR from large RNA fraction of sample used for small RNA library preparation. (B) Mapping of siRNAs (21 nt) to SINV genome from SINV infected Aag2 cells treated with control (dsFluc), Ago2, Ago3 or Piwi4[O1] dsRNA. (C) Mapping of piRNAs (24-30 nt) to SINV genome from SINV infected Aag2 cells treated with control (dsFluc), Ago2, Ago3 or Piwi4 dsRNA. The number of small RNAs is normalized to the number of SINV genome copies for each dsRNA treatment. (D) SINV genomic RNA measured by RT-qPCR from large RNA fraction of sample used for small RNA library preparation. (E) Size distribution plot of small RNA mapping to transposons from infected Aag2 cell treated with control, Ago2, Ago3, or Piwi4 dsRNA. (F) Size distribution plot of beta-elimination resistant small RNA mapping to transposon from infected Aag2 cells treated with control, Piwi4, or Ago3 dsRNA.