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. Author manuscript; available in PMC: 2016 Jun 23.
Published in final edited form as: Cell Rep. 2015 Jun 11;11(11):1714–1726. doi: 10.1016/j.celrep.2015.05.032

Figure 3. Constructing a library of siRNAs targeting virus or IFN induced genes.

Figure 3

(A) Schematic depicting the structure of pre- and mature miRNA duplex with the 5p (orange) and 3p (blue) strands highlighted. Beneath this is a heat map of relative read counts derived from small RNAseq analysis from small RNAs derived from A549s infected with the IAV library. Results are shown as the proportion of the 5p and 3p produced for each siRNA.

(B) Quantitative RT-PCR (qPCR) of mRNA derived from mouse embryonic fibroblasts mock treated or infected (MOI=3) with viruses targeting IRF7, CXCL10, RIG-I, OASL, IFIT3, and IFIT1. Each qPCR was generated to determine the expression profile of the cognate target. Each virus set included two unique siRNA hairpins (−1 or −2). Error bars, SD, *p<0.05.

(C) RNA and protein analysis of ISG15 from MEFs mock treated or infected (MOI=3) with the corresponding siISG15 viruses (Isg15-1 or Isg15-2). Top panel depict qPCR analysis for ISG15 as described in (B), bottom panel depicts a western blot of whole cell extract derived from 293 cells transfected with epitope tagged (V5) murine ISG15 and infected with the aforementioned viruses. Immunoblots were probed form V5, IAV NP, and Actin.