Pupa and adult flies were fixed in Methacarn (6:3:1 of methanol:chloroform:acetic acid) to preserve tissue integrity and nucleic acids prior to dissection.  Indirect flight muscles (IFMs) (inclusive of dorsal longitudinal muscles (DLMs) and dorsal ventral muscles (DVMs)) were microdissected separately from ventral nerve cord (VNC) (separated from central brain). A total of three replicates of n = 50 for each tissue/treatment/stage (n=150 total) were collected. Dissected tissues were immediately homogenized and preserved in TRIzol™ Reagent (Thermo-Fisher Scientific) and frozen at -80°C until processed for total RNA extraction. RNA liibraries for RNA-Seq were prepared using TruSeq Stranded Total RNA Gold as per manufacturers protocol.