design: The total RNA was isolated from whole brain using a Zymo RNA Miniprep extraction kit. Poly(A) RNA was isolated using the NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490) according to the manufacturers protocol with one adjustment: 60 L magnetic oligo d(T)25 beads were used for the initial binding to accommodate the increased total RNA input. Immediately following isolation, library preparation from poly(A) RNA was performed using the Direct RNA Sequencing Kit (SQK-RNA002) and the manufacturers protocol (version DRS_9080_v2_revM_14Aug2019). One Nanopore flow cell (R9) was used for each sample. Flow cell check was performed to confirm 800 active pores immediately prior to using each flow cell, following manufacturers protocol (Flow Cell Check protocol version PQE_1004_v1_revAC_06Jan2016). Flow cell priming was performed according to the manufacturers protocol for direct RNA sequencing. Data were acquired in minKNOW UI version 4.1.22 using default settings. Flow cells were run to depletion of the RNA input, approximately 25 hours.