EED Antibody - #DF7308

Figure 6. CXCL1exo-dead 520 transcriptionally increases PD-L1 expression in macrophages by activating EED signaling. (A) The mRNA level of PD-L1 in Raw264.7 macrophages was significantly increased by 10 ng/ml murine CXCL1 and 50 μg/ml exo-dead treatment for 24 h; n = 3. (B) The promoter activity of PD-L1 in Raw264.7 cells when treated as indicated for 24 h; CXCL1-NA concentration: 5 μg/ml; n = 3.（C) Diagram of the DNA-pull down-MS assay. The DEPs that bind with the PD-L1 promoter region of Raw264.7 macrophages after CXCL1 treatment for 24 h were analyzed by mass spectrometry. The TFs of PD-L1 were predicted using the hTFtarget database. EED was selected as the potential transcription factor by taking the intersection of the DEP set and the TF set. (D–E) The expression levels of EED and PD-L1 in Raw264.7 cells when treated as indicated for 48 h. Murine CXCL1 concentration: 10 ng/ml. Scale bar: 5 μm. (F) The combinational effect of CXCL1 treatment and EED knockdown on the promoter activity of PD-L1 in Raw264.7 cells when treated as indicated for 24 h; n = 3. (G) The binding activity of EED with the promoter fragment of PD-L1 in Raw264.7 cells when treated as indicated for 48 h was investigated by CHIP-PCR assay. (H) The antisense mutation of the EED binding region in the PD-L1 promoter significantly abrogated the induction effect of CXCL1 on PD-L1 promoter activity in Raw264.7 macrophages