EC_tagging pulse-chase to measure mRNA decay in Drosophila larval neuroblasts and neurons

Nascent RNA was tagged with EU, via EC feeding, in targeted cells (neuroblasts or neurons) then purified after the initial pulse labeling or after a chase using media containing excess unmodified uridine Neuroblasts were targeted using insc-GAL4 to drive UAS-CD:UPRT. Neurons were targeted using nSyb-GAL4 to drive UAS-CD:UPRT. Tagged RNA was purified following a 12 hour pulse feeding of 5-ethynylcytosine or at subsequent chase timepoints in which larvae were fed an excess of unmodified uridine: 3 hour chase, 6 hour chase, 12 hour chase.