KAP1–mediated control of a ERE-based regulatory element modulates the lineage-specific expression of TLR1 (A) UCSC Genome Browser view of TLR1 locus. Displayed tracks include RNA-seq for control (shLUC) and KAP1 knockdown (shKAP1) activated CD4 + T cells; RefSeq genes and retroelement annotation (modified from UCSC Genome Browser as described in the Methods); DNase cluster (ENCODE); KAP1 and H3K9me3 ChIP-seq tracks for resting and activated CD4 + T cells; public H3K9me3, H3K4me3, H3K4me1, and H3K27ac ChIP-seq tracks for Th1 activated CD4 + T cells and/or mature neutrophils; primers used in (B) located at varying distances relative to the TSS. The dashed box highlights the KAP1-bound region. The repetitive elements on which KAP1 binding is centered are shown in red. (B) ChIP-qPCR analysis of H3K9me3, H3K4me1 and H3K27ac around KAP1 binding site at the TLR1 locus in control and KAP1 knockdown cells. Values are normalized to their respective total inputs, to the total H3 protein levels and to GAPDH . (C) RT-qPCR analysis of KAP1 and TLR1 mRNA expression in activated CD4 + T lymphocytes transduced with shLUC or shKAP1-expressing LVs. (D) RT-qPCR analysis of TLR1 mRNA expression in activated CD4 + T lymphocytes and in the human promyelocytic leukemia cell line HL-60. HL-60 cells were differentiated into neutrophil-like cells in culture medium in the presence of 1.3% DMSO for five days. (E) RT-qPCR analysis of KAP1 and TLR1 mRNA expression in untreated or DMSO-treated HL60 cells, transduced with shLUC or shKAP1-expressing LVs. Error bars represent SEM of biological replicates (n=3), *p
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