splenocytes mechanically dissociated and flow sorted for CD8+ T cells binding to the ovalbumin SIINFEKL tetramer Gene expression, feature barcoding and TCR sequencing libraries were prepared using the 10x Genomics Single Cell 5' Reagent Kits v2 (Dual Index) following manufacturer user guide (CG000330 Rev B). The final libraries were diluted to ~10nM for storage. The 10nM library was denatured and further diluted prior to loading on the NovaSeq6000 sequencing platform (Illumina, v1.5 chemistry, 28bp/98bp paired end for gene expression and feature barcoding, 150bp paired end for TCR libraries). 24h samples are composed of a combined three biological replicates in equal proportion, and sequenced as four technical replicates for naïve and listeria-infected d9 samples were hashed using BioLegend TotalSeq™-C03 series antibodies, Hashtags 1-4 for each biological replicate