Fig. 4: KDM2A upregulates PD-L1 expression in mammary fibroblasts via transcriptional activation.
From: Lysine demethylase 2A expression in cancer-associated fibroblasts promotes breast tumour growth
a Expression of PD-L1 mRNA in RMF-EG and K2A-1 cells was investigated by qRT-PCR. b Flow-cytometry analysis was done to study the cell surface PD-L1 in RMF-EG and K2A-1 cells. In addition, the percentage of PD-L1-positive cells was compared. c RMF-EG cells were co-cultured without or with MDA-MB-231 cells and incubated without or with daminozide for 24 h. The percentage of PD-L1-positive cells was compared in different experimental groups. In addition, RMF-EG and KDM2A-depleted RshKDM2A cells were co-cultured with MDA-MB-231 cells. After 24 h, PD-L1-positive cells were determined. d RMF-EG cells were pretreated without or with daminozide and then stimulated with IL-6 (20 ng/ml) for 24 h. Expression of PD-L1 was studied by qRT-PCR. e Genomic DNA was harvested from RMF-EG and K2A-1 cells and ChIP assays were performed by using various antibodies to investigate the binding of KDM2A to human PD-L1 gene promoter and the methylation status of H3K36 and H3K4 in the promoter. The genomic region amplified by the PCR primers was shown. *P < 0.05; **P < 0.01; ***P < 0.001.
