Project description:CD8+ DCs play key role in CD8+ T cell priming, however, the underlying signaling mechansim is unclear. We used a data-driven network-based systems biology approach and identified Hippo signaling kinases as key selective modualtors in CD8+ DCs. We focused on Mst1/Stk4 to further investigate the novel function of Hippo signaling in CD8+ DCs. All transcriptional profies were evalated by microarray. 1) We used microarrays of total DCs in previous (GSE98481) and current studies to reverse engineer a DC-specific signaling interactome (DCI). 2) We integrated DCI with profiles of sorted CD8+ with CD8- DCs to identify signaling drivers of CD8+ DCs. 3) We profiled and compared Mst1/2-KO with WT cells from total, CD8+ and CD8- DCs to understand the mechanism of Mst1 in CD8+ DCs.
Project description:In order to explore the regulatory mechanism of MST1 kinase on Tfh differentiation and function, we sorted Tfh cells in the spleen of WT and Mst1 KO mice 8 days after intraperitoneal sensitization, and extracted total RNA from the collected cells as samples for sequencing analysis.
Project description:To comprehensively understand how dendritic cells (DCs) are reprogrammed by lung fibroblasts- and their derived COX-2/PGE2, we employed lung fibroblasts isolated from WT or Ptgs2-/- mice, and collect their conditioned medium (CM) to stimulate the ex vivo cultured bone marrow (BM)-derived DCs (BM-DCs), with the PGE2 treatment as a control. After the treatment, BM-DCs were harvested for RNA extraction and the transcriptional profiles were analyzed by RNA sequencing (RNA-seq).
Project description:Analysis of the specific transcriptional changes on DCs provided by direct pattern recogition receptor (PRR) or IFNAR signaling that are required for DC maturation after poly IC stimulation. Results provide important information about the intricate differentiation process of DC maturation and the importance of type I IFNs for DC immunogenicity. WT/IFNA-/- or WT/PRR-/- mixed-chimera mice were injected with 50 ug Poly-IC i.p. in vivo 4 and 14 hr later, wild type and KO CD11chi CD3- DX5- B220- DCs were FACS sorted based on CD45.2 expression. Total RNA was isolated and expression profile was compared between unstimulated and activated WT and KO DCs.