Skip to main content
. 2020 Jul 29;8:622. doi: 10.3389/fcell.2020.00622

FIGURE 1.

FIGURE 1

Acheron expression during ISM development. (A) RNA-seq analysis of Acheron mRNA in the ISMs from day 13 until day 18, the day of adult eclosion. “20E” represents animals that were injected on day 17 to delay the time of cell death and then assayed on day 18. Acheron expression changed during development and was induced on day 18 (p = 0.002) but blocked by pre-treatment with 20E (p = 0.025, n = 3 independent replicates for each stage of development). (B) qPCR quantification of Acheron mRNA late in development and following 20E treatment. Only the day 18 sample was significantly different from day 13 (p = 0.02). (It should be noted that these data agree well with our independent validations of this RNA-seq dataset using qPCR; Tsuji et al., 2020). (C) (Left) Western blot of Acheron expression at carefully timed stages between day 15 of pupal/adult development and 2 h post-eclosion (PE). Acheron expression was prevented by treatment with 20E. The blots were also probed for tubulin (bottom), which served as a loading control. (D) Detailed examination of Acheron expression on late day 18 starting at 4:30 p.m. Animals in the colony eclose at about 5:30 p.m. (E,F) Immunohistochemical staining of Acheron before (E) and after (F) adult eclosion.