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. 2016 Dec 10;6:221–232. doi: 10.1016/j.omtn.2016.11.007

Figure 2.

Figure 2

RRV-U6-miRGFP Has Higher GFP Downregulation Activity than RRV-U6-shGFP and Does Not Trigger IFN Response

(A) RRVs produced from transient transfection in 293T were used to infect U87-MG/GFP cells at a MOI of 0.1 (see Figure S1 for MOI of 1 and 10). The GFP downregulation activity was evaluated and quantified by flow cytometric analysis for MFI. Both uninfected cells and cells infected with RRV-yC2 were included as a positive control. The percentage of GFP downregulation is relative to the uninfected cells. The data shown represent mean ± SD of triplicates performed from one of multiple independent experiments. (B) Vector stability of RRV-U6-shGFP and RRV-U6-miRGFP using genomic DNA from infected U87-MG/GFP cells at 30 days post infection was analyzed by endpoint PCR. The DNA molecular marker (1 Kb Plus marker, Invitrogen) is included in the first lane. The right and left arrows indicate the expected size of the PCR products for the RRV-shGFP (514 bp) and for the RRV-miRGFP (844 bp), respectively. (C) Induction of OAS1 gene expression induced by IFN response in U87-MG cells infected with RRV-U6-shGFP or RRV-U6-miRGFP at a MOI of 0.1, 1, and 10. U87-MG cells treated with exogenous recombinant human IFNα were included as positive control. The values are presented as the means ± SD of triplicates.