Quantification of MCMV and CD70 in DCs

Spleen DNA was prepared and analyzed for MCMV genomes using quantitative (q)PCR as previously described (49 (link)). For analysis of Cd70 expression, splenic CD11c+ DCs were positively selected using CD11c+ MACS microbeads (Miltenyi Biotec, San Diego, CA). RNA was isolated using TRIzol (ThermoFisher Scientific) according to manufacturer guidelines, and converted to cDNA using Advantage RT for PCR Kit (Clontech, Mountain View, CA). Cd70 cDNA was amplified using gene-specific primers: Cd70-Forward, 5′-TGC TGT TGG TTT CAT TGT AGC G-3′; Cd70-Reverse, 5′-ATC CTG GAG TTG TGG TCA AGG G-3′, as reported (50 (link)). Hprt was also amplified using gene-specific primers (22 (link)) and used to normalize and compare Cd70 expression in infected and naïve DCs.