Neuronal expression of bromodomain proteins alter neuronal health and peripheral protein homeostasis to promote longevity
Neuronal expression of bromodomain proteins alter neuronal health and peripheral protein homeostasis to promote longevity
For all day 1 RNA-seq experiments, after bleaching and L1 arresting, worms were grown at 22°C for 72 hours (i.e., 3 days) to obtain day 1 adult animals. For day 7 RNA-seq experiments, we used N2 worms, as animals no longer have germline at day 7 of adulthood. After bleaching and L1 arresting, animals were grown at 20 °C for 72 hours (i.e., 3 days). Animals were then transferred to FUDR and grown for an additional 6 days prior to sample collection. Roughly 1,000 worms per condition were harvested with M9, transferred to 1 mL of TRIzol Reagent.  After transfering to 1 mL of TRIzol Reagent, animals are subjected to three freeze/thaw cycles between liquid nitrogen and a 37°C bead bath, with 30 s vortexing between cycles. Following the final thaw, chloroform was added at a 1:5 ratio (chloroform:TRIzol), and RNA was separated by centrifugation in heavy gel phase‐lock tubes (VWR, 10847‐802). The aqueous phase was combined 1:1 with isopropanol, and purification was performed with a QuantaBio Extracta Plus RNA kit (95214) per the manufacturer's instructions. Library construction was performed by Novogene using their standard pipeline.