Overview of the high-resolution RNA isoform annotation in adult female P. puparum. (A) The workflow for sequencing and analysis, which includes multiple sequencing strategies such as PacBio HiFi Iso-Seq, stranded RNA-seq, CAGE-Seq, and PAS-Seq, as well as bioinformatic analyses for full-length transcriptome reconstruction and error corrections. (B) Distribution of the number of isoforms per gene. (C) Pie charts illustrating the composition of coding and noncoding isoforms/genes. (D) An example gene (longitudinals lacking gene, lola) with 266 isoforms. The top panel shows CAGE reads, the middle panel shows PAS reads, and the bottom panel shows a subset of isoforms (for complete set, refer to Supplemental Fig. S4). (rpm) reads per million. (E) An example demonstrating the precise annotation of two overlapping genes from different strands. The top panel shows strand-specific RNA-seq (blue represents Watson strand mapping reads; red represents Crick strand mapping reads), and the bottom panel shows isoforms. (VG) venom gland, (CA) carcass, (rpm) reads per million.
