Hypothesis testing of major transitions during diapause
In order to gain additional insights beyond our GSEA, we next identified
differentially expressed miRNAs during the period when pupae
transitioned from the deepest portion of diapause to post diapause
quiescence (day 24-144). During this period butterflies were held at
2°C, allowing us to identify miRNAs associated with diapause progression
independent of temperature change. There were eight DEGs identified in
both the head and abdomen tissue, and of those, only two were
differentially expressed in both tissues, miR-14-5p and miR-2a-3p. These
two miRNAs have inverse normalized expression patterns, where miR-14-5p
increases in expression early in diapause, and declines to a minimum
expression after 114 days (Figure 6). miR-2a-3p has an expression
pattern that starts off at the lowest expression, and increased through
time until day 114, where it stays until 144, afterwards it starts to
decline (Figure 6).