Figure 3.

Activation of the TLR9-MyD88 signaling pathway and a distinct transcriptomic signature of GzmA-stimulated pDCs. (A) IFN-α production by FL-DCs from WT or several types of KO mice stimulated with GzmA. FL-DCs were cultured in the presence of GzmA for 24 h and the supernatants were measured by ELISA for IFN-α (mean ± SEM, n = 4) medium vs. GzmA, ^**p < 0.01, ^***p < 0.001 (Mann-Whitney). (B) Analysis of differential gene expression in pDCs before and after stimulation with GzmA. pDCs unstimulated or stimulated with GzmA for 8 h were sorted. Differential genes associated with IRF signaling, TLR signaling, IFN signaling, MyD88 signaling, and interferon-responsive genes in pDCs were assessed by RNA-Seq. The gene expression analysis was performed using an R based package, EdgeR with the raw counts from RNA-seq data. Then, expression levels (expressed as FPKM) from selected genes of pDCs under these two conditions were compared (n = 3, mean ± SEM). ^*p < 0.05, ^**p < 0.01 (paired Student's t-test).