YOD1-C160S expressed in BMDCs has no effect on direct antigen presentation. (A) Schematic of direct antigen presentation experiments. Expression of YOD1-C160S in BMDCs was induced by adding 1 μg/mL of doxycycline to the culture. BMDCs were either pulsed with the class I restricted peptides SIINFEKL or KNYIFEEKL, or infected with recombinant MHV-68 virus expressing SIINFEKL. BMDCs were then cocultured with CFSE labeled OT-I or ORF8 TN cells. Activation, proliferation, and cytokine production by T cells was analyzed. (B-C) FACS plots and bar diagram show the activation of OT-I cells at different SIINFEKL concentration as indicated by CD69 surface expression. (D) FACS plots show the proliferation of OT-I Tg (top panel) or ORF8 TN cells (bottom panel) when cocultured with the respective peptide-pulsed BMDCs. (E) IL-2 levels in the culture supernatants as measured by ELISA are shown. (F-G) Proliferation (F) and IL-2 production (G) by ORF8 TN and OT-I cells are shown. All experiments were repeated 3 times and data are represented as mean ± SEM.