FIG 4.

Quantification of mRNAs synthesized by ΔC7K1 and WT virus and translation of transfected mRNAs. (A) mRNA quantification. HeLa and SAMD9^−/− cells were infected in triplicate with ΔC7K1 or WT virus at a multiplicity of infection of 3 PFU/cell, and RNA was extracted at 2 and 8 h, reversed transcribed, and quantified by ddPCR using primers specific for I3, D13, and A3 transcripts and rRNA. The absolute amounts of viral mRNA in each sample were calculated using the 18S rRNA concentration and the number of ribosomes in HeLa cells. (B) Translation of mRNA synthesized in vitro. Control HeLa and SAMD9^−/− cells were infected in triplicate with ΔC7K1 at a multiplicity of infection of 3 PFU/cell in the absence or presence of AraC and then transfected with capped, methylated, and polyadenylated mRNA with a 5′ poly(A) or non-poly(A) leader preceding the FLuc ORF. FLuc was measured after 6 h. Error bars represent SEM.