Neuromodulation can be simple: myoinhibitory peptide, contained in dedicated regulatory pathways, is the only neurally-mediated peptide modulator of stick insect leg muscle
Legs were dissected as described above. RNA was extracted from the ExtTi and FlxTi muscles of a single middle leg using TRIzol (Thermo Fisher Scientific) following manufacturer recommendations. RNA quality was estimated using RNA concentration (ng/Î¼L) and RNA integrity number as implemented in Agilent 2100 Bioanalyzer. Libraries were prepared using 1 Î¼g total RNA with the IlluminaÂ® TruSeqÂ® stranded RNA sample preparation kit. PolyA containing mRNA were purified using oligo-dT attached magnetic beads and then fragmented and primed for cDNA synthesis. Fragmented RNA was reverse transcribed using random hexamers to produce first strand cDNA; second strand cDNA was synthesized with DNA polymerase I and RNase H. After end repair and A-tailing, multiple indexing adapters for hybridization onto a flow cell were ligated. Products were enriched with PCR and purified to create the final cDNA library. Sequencing was done on an Illumina HiSeq 4000 sequencer with 75 paired ends at Cologne Center for Genomics (CCG, Cologne, Germany).