Segment 6 coding strategies. (A) RNA extracts obtained from HEK 293 cells infected with Upolu virus (UPOV) or Jos virus (JOSV) (cellular RNA [cR]) or from DNase- and RNase-treated supernatants (genomic RNA [gR]). cDNA was amplified with primers located upstream of a potential splice region (p1) and downstream at an mRNA polyadenylation signal (p2) or at the segment terminal sequence (p3). M indicates molecular size markers. Only a single-size amplification product was observed with the UPOV template (bands 1 and 2), whereas differently sized products were generated with the JOSV template (bands 3 to 5). (B) Relevant sequences obtained from the respective bands shown in panel A. (C) Schematic of segment 6 coding strategies of Thogoto virus (THOV) and JOSV, and of Dhori virus (DHOV), Aransas Bay virus (ABV), and UPOV, indicating locations of primers p1, p2, and p3; ML or M ORF termination codons (ochre, opal, and amber); and splice sites.