GSM5135523: WT DTx + aPD-L1 (C1_2); Mus musculus; RNA-Seq
Mice were sacrificed and spleen isolated. Single cell suspnesion was created by grinding spleens though a filter. Afterwards, red blood cells were lysed with ACK-buffer for 4min. Splenocytes were staining with antibodies for 30min at 4 degrees and sorted for the desired population using FACS AriaIII. CD8+, PD-1+, TIM-3 negative single-cells were sorted from the spleen of chronically infected mice using a FACSAria III (BD Biosciences), before being encapsulated into droplets with the ChromiumTM Controller (10x Genomics) and processed following manufacturer's specifications. Transcripts captured in all the cells encapsulated with a bead were uniquely barcoded using a combination of a 16 bp 10x Barcode and a 10 bp unique molecular identifier (UMI). cDNA libraries were generated using the Chromium™ Single Cell 3' Library & Gel Bead Kit v2 for the WT untreated sample or v3 for WT and XCR1-DTR samples treated with aPD-L1 and DTx (10x Genomics) following the detailed protocol provided by the manufacturer.