Figure 1. Experimental design used to investigate the effect of
acclimation at 36 °C vs. control treatments at 25 °C in parents (P) and
F1 offspring. Acclimation treatments in the parental
generations were performed on adult stages, while offspring acclimation
was performed on either larva or adult samples. All samples for RNA-Seq
were collected in the offspring generation as a result of the four
different combinations of acclimation treatments. The parental effect
was used to assess transcriptional transgenerational plasticity (TGP),
while the offspring acclimation was used to estimate within-generation
plasticity (WGP).
We sequenced nearly 400 million paired-end Illumina reads across the 24
RNA-seq libraries. Of these, an average of 17 million reads per library
mapped to the reference genome following trimming and filtering
of sequence reads. An independent read-count matrix for each gene
feature was generated (i.e., exons, junctions, introns, and
gene-wide). All relative changes of transcriptional plasticity were
estimated by comparing read counts between acclimated samples at 36 °C
and control samples at 25 °C as performed in the parental or offspring
generations (Figure 1).