MINOR inhibition in DCs improved cytokine expression and T-cell expansion. BMDCs were transduced with either LV-control-siRNA or LV-MINOR-siRNA, pulsed with MHC II–restricted HA peptide and analyzed for their ability to secrete cytokines in vitro and stimulate CD4⁺ T cells in vivo. RNA was isolated from BMDCs and analyzed for IL-12 expression by quantitative PCR (A). For in vivo analysis, anti-HA T cells (6.5 cells) were injected 1 day before DC vaccination with transduced DCs; T-cell expansion was measured by harvesting spleens and measuring percentage of clonotypic 6.5 cells by FACS. (B) Percentages of clonotypic cells, from mice vaccinated with LV-control-siRNA– or LV-MINOR-siRNA–transduced DCs (or unvaccinated control, phosphate-buffered saline) pulsed with MHC II–restricted HA peptide. (C) Statistical analysis of percentages of cells expressing TNF-α by 6.5 CD4⁺ T cells rechallenged with HA-specific peptides. Results from 2 separate experiments are shown; statistical analysis is by t test.