RNA sequencing of murine bone marrow cell-derived cDC1 treated with PGE2

RNA sequencing was used to characterize PGE2-mediated changes in the gene expression profile of conventional type 1 dendritic cells (cDC1). Our analysis shows that treatment of cDC1 with PGE2 or conditioned medium from PGE2-producing tumors induces transcriptional changes in resting cDC1. cDC1 activated with a TLR3 ligand after PGE2 pre-treatment show alterations in the expression of activation induced genes. Overall design: cDC1 were differentiated from C57BL/6J bone marrow cells in vitro over 14 days using FLT-3 ligand and GM-CSF. CD103+ cDC1 were sort-purified and treated for 4h with PGE2 (100ng/ml) or conditioned medium from PGE2-producing BRAFV600E melanoma cells or PGE2-deficient Ptgs1/Ptgs2-/- BRAFV600E melanoma cells. In some groups, antagonists for the PGE2 receptors EP1, EP2, EP3 or EP4 were added to the culture. For analyses of changes upon cDC1 activation, cDC1 were treated for 24h with PGE2 and stimulated for the last 4h with the TLR3 ligand poly I:C.