GSM8907886: lung, post-inoculation Day28-1; Mus musculus; RNA-Seq
For CD45+ immune cells isolation, the Miltenyi MACS bead sorting system (Miltenyi Biotec,130-052-301) was used according to the manufacturer's guidelines. After isolation, cell number and viability were determined with Trypan blue using a Countess II Automated Cell Counter (Thermo Fisher). The collected cells were then resuspended at 1×105 cells/mL with a final viability of > 90%, assessed by the Countess Cellular suspension was loaded on a 10X Genomics instrument for Gel Bead-In-EMlusion (GEMs) generation and barcoding according to the manufacturer's protocol. The GEMs were then mixed with enzymes to produce full-length, barcoded cDNAs that were then amplified by PCR to generate sufficient mass for library construction. To prepare the library construction, amplified cDNA was enzymatically fragmented into 200bp~300bp fragments, which were then end-repaired, polyA-tagged and amplified by PCR for final libraries. Samples were sequenced on an Illumina NovaSeq platform.