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. 2022 Jan 20;50(3):1734–1752. doi: 10.1093/nar/gkac022

Figure 6.

Figure 6.

Nascent RNA silencing is mediated by Ago2 cleavage. (A) Images of northern blots showing signals of the tested RNA substrate, which was derived from the genomic sequences of SPINT1 containing its predicted target site in intron 2 and flanking sequences, and the resulting cleaved RNA at different time points after the addition of components indicated above each image. The experimental strategy is illustrated in the diagram on top. (B) Bar chart showing the RNA levels of SPINT1 intron substrate containing tsRNA target site, in presence or absence of recombinant Ago2 and tsRNA targeting SPINT1. (C) As in (B), in presence of Flag tagged wt and Ago2D669A mutant. (D) Bar chart showing the fold change of mouse nascent Egfr transcripts, measured by qRT-PCR, upon mock transfection and transfection of plasmids expressing wild type human Ago2 (wt hAgo2) and mutant form of human Ago2 D669A (hAgo2D669A) in MEFs. (E) Bar chart showing mouse nascent EGFR levels in wt MEF Ago2-/- transfected with hAgo2, siRNA targeting Dicer and tsRNA targeting human EGFR.