GSM7429846: mixed sample of medulloblastoma, CT-179-treated, POx-resiquimod-treated, untreated control; Mus musculus; RNA-Seq
Brains were freshly harvested from treated and control mice, and then cut in half sagittally. One half was then drop-fixed in 4% paraformaldehyde for IHC, and tumor was dissected free from the other half. Dissected tumors were then processed for split-seq analysis using the Parse WT 1.0 kit. For processing, tumors were dissociated using the Papain Dissociation System (Worthington Biochemical) according to the manufacturer's protocol, as in our prior studies, https://www.nature.com/articles/s41467-019-13657-6#Sec18. Cells were then counted and fixed using the Parse WT 1.0 Fixation Kit, per manufacturer's instructions. For library construction, fixed cells were plated out in the Parse WT 1.0 bar coding kit, according to manufacturer's protocol. A total of 8 sublibraries were generated and validated by Tapestation. Sequencing was then performed on an estimated 60,000 total cells across the three conditions.