Figure 1

Gene selection process by metabolic pathway. Our starting point was the database with 55,000 annotated genes from the R. arenarum transcriptome previously assembled by us¹³. The first step was a hypothesis-driven selection of annotated genes from five pathways recognized as affected by organophosphorus pesticides at enzyme activity or metabolic product levels: PM polyamine metabolism, AS antioxidant system, DS detoxifying systems, TF transcription factors, signaling pathways; and Housekeeping (HK) genes tested in amphibians. The next step was checking the selected transcript sequences and their predicted amino acid sequence identities with the annotated genes by BLASTN and BLASTP, from which about 94% of isoforms were confirmed (light blue vs. brown slices). The last step was checking the transcription levels for each isoform to discard erratic low values, using the differential expression data corrected by the trimmed mean of M-values (TMM) normalization method, accepting 76% of them (light green vs. yellow slices). Note that, on average, there were 2 accepted transcripts per annotated gene.