Experimental design and RNA-Seq
Previous studies have demonstrated adaptive within- (WGP) and transgenerational plasticity (TGP) of heat tolerance following acclimation treatments at 36 °C in parents and offspring of D. mojavensis (Diaz et al., 2020). Here, we investigate the underlying transcriptional bases of these types of plasticity by using the same experimental design, which allows us to address fundamental questions on how selection drives adaptive WGP and TGP while simultaneously considering larval and adult stages of D. mojavensis . Our approach involves a full factorial design where parents and offspring were exposed to either an acclimation temperature of 36 °C or a control treatment at 25 °C (Figure 1). RNA-Seq libraries were sequenced from larva and adult samples collected in the offspring generation, with three biological replicates for each of the four combinations of acclimation treatments (Figure 1).