Total RNA was isolated from cell lysates of brain tricultures and IMHBOs using the Qiagen RNeasy Mini kit following manufacturer's instructions. RNA samples were quantified using Qubit 4.0 Fluorometer (Life Technologies) and RNA integrity was checked with RNA Kit on Agilent 5300 Fragment Analyzer (Agilent Technologies). RNA sequencing was performed by Azenta Life Sciences/Genewiz (Germany). Briefly, libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina following manufacturer's instructions (NEB). Sequencing libraries were validated using the NGS Kit on the Agilent 5300 Fragment Analyzer (Agilent Technologies) and quantified by using Qubit 4.0 Fluorometer (Invitrogen). The sequencing libraries were multiplexed and loaded on the flowcell on the Illumina NovaSeq 6000 instrument according to manufacturer's instructions. The samples were sequenced using a 2x150 Pair-End (PE) configuration v1.5. Image analysis and base calling were conducted by the NovaSeq Control Software v1.7 on the NovaSeq instrument.