Setd1a and Setd1b loss-of-function in murine macrophages

Setd1bKO primary murine bone marrow-derived macrophages (BMDMs) were treated with lipopolysaccharide (LPS) or dexamethasone and LPS (Dex+LPS) and gene expression differences in response to treatment analysed by PolyA RNA-Sequencing. No Dex-treatment dependent gene expression differences were identified. Setd1aDel/+ Raw264.7 cells with reduced Setd1a expression were analyzed with regards to their reponse to Dex when inflammatorily challenged with LPS by mRNA-Seq. We observed reduced GR-dependent gene acivation in Setd1a hypermorphic Raw264.7 cells. Wild type and Setd1aDel/+ Raw264.7 cells were treated with LPS or LPS and interferon beta (IFNB1) to show the IFNB1-dependent loss of gene expression in LPS-stimulated Setd1aDel/+ cells. We profiled mRNA expression by RNA-Seq in form wild-type or Setd1bKO BMDMs treated with either vehicle (0.1% EtOH) for 16h, 6h LPS (0.1% EtOH, 100ng/ml LPS) or 16h Dex+6h LPS (1 uM Dex, 100ng/ml LPS) in order to study the function of Setd1b as GR?s nuclear interaction parner. For the analysis of Setd1a-dependent gene expression, we performed mRNA-Seq from wild-type or Setd1aDel/+ Raw264.7 cells. Raw264.7 cells were treated with Dex for either 16h (vehicle: 16h 0.1% EtOH; LPS: 6h 100 ng/ul LPS, Dex+LPS: 16h 1 uM Dex, 6h 100 ng/ul LPS) or 6h (vehicle: 6h 0.1% EtOH; LPS: 6h 100 ng/ul LPS, Dex+LPS: 6h 1 uM Dex, 6h 100 ng/ul LPS; Dex: 6h 1 uM Dex). Furthermore, we profiled wild type and Setd1aDel/+ RAW264.7 cells after 6h of LPS stimulation (100 ng/ml) and compared the effect of Setd1a deletion on gene expression to LPS+IFNB1 (6h 100ng/ml LPS + 10 ng/ml IFNB1) stimulated cells of both genotypes. Three biological replicates were sequenced per treatment and genotype.