RNA-seq Analysis of the 5-HT2B Receptor Agonist BW723C86 on OGD-induced HT22 Cells in a Transwell Co-culture System with BMDMs

To investigate the transcriptional changes underlying this crosstalk, we employed a Transwell co-culture system. Specifically, Bone marrow-derived macrophages（ BMDMs) were co-cultured with HT22 mouse hippocampal neurons subjected to 2 hours of oxygen-glucose deprivation (OGD) to model ischemic conditions.

This setup allowed for the paracrine exchange of signals without direct cell contact. Following OGD, the co-culture was treated with either the 5-HT2B receptor agonist BW723C86 or a DMSO vehicle control. We then performed bulk RNA-sequencing on the microglial/BMDM population to profile their transcriptomic response to neuronal injury signals and receptor agonism.