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. 2022 Dec 3;14(12):2711. doi: 10.3390/v14122711

Figure 4.

Figure 4

Analysis of the replication of wt FHV RNA1 and its mutant derivative RNA1-noB2 replication in insect and mammalian cells. (A) Schematic maps of wildtype RNA1 and an RNA1-derivative that lacks coding potential for B2 due to mutations of initiator and downstream methionine codons that are silent in the -1 frame encoding protein A sequence. (B) B2 is a strong suppressor of RNA-interference (RNAi), and RNA1-noB2 does not support stable accumulation of RNA1 and subgenomic RNA3 and protein A in Drosophila S2 cells that have a strong antiviral RNAi response (left lanes). By contrast, RNA1-noB2 launches RNA1 and RNA3 replication levels very similar to wildtype RNA1 in mammalian BSRT7/5 cells that do not display robust RNAi action (right lanes). (C) Fluorescence images of BSRT7/5 cells confirm accordingly that B2 is not needed and show that protein A and dsRNA replication intermediates are observed at the mitochondria of wildtype RNA1 and RNA1-noB2 transfected cells at similar levels and distribution.