We next determined the highest 20 expressed genes detected across treatments and found the “INTACTlabel” (manually added to the Dm6 transcriptome) was highly expressed across the libraries, demonstrating the robust nature of the GAL4/UAS binary expression system and further validating the specificity of the MB isolation technique. Most of the highly transcribed genes were RNA-associated genes (Figure S3Ci), which we posit to be a mix of transcripts from rough endoplasmic reticulum that was pulled down during nuclear isolation, as well as newly transcribed or nuclear-localized RNAs. We further curated the highest 20 mean expressed genes by excluding RNA-associated genes (Figure 1B). Together, these descriptive analyses indicate that the libraries represent comparable MB data sets in which lasting experience-dependent molecular effects could be explored.
