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. 2020 Oct 7;124(2):484–493. doi: 10.1038/s41416-020-01112-z

Fig. 2. Cancer-derived cytokines upregulate KDM2A expression in normal fibroblasts and transform normal fibroblasts into CAFs.

Fig. 2

a Co-culture of normal human mammary fibroblasts (RMF-EG) with MCF-7 or MDA-MB-231 (231) breast cancer cells for 48 h and the expressions of KDM2A in RMF-EG cells were investigated by western blotting. b Normal RMF-EG mammary fibroblasts were treated with various doses of IL-6 and the KDM2A protein level was studied. c Normal RMF-EG mammary fibroblasts were treated with various doses of TNF-α and the KDM2A protein level was investigated. d RMF-EG cells were transfected with pEZX-PG04-KDM2A gene promoter construct and treated with recombinant IL-6 or TNF-α (20 ng/ml) for 24 h. The promoter activity was determined by using the Secrete-Pair Dual Luminescence Assay Kit. *P < 0.05. e Normal RMF-EG mammary fibroblasts were transfected with KDM2A expression vector and two independent stable clones K2A-1 and K2A-2 were established by antibiotics selection. Expression of KDM2A, FAP and PDGFR-α was investigated by western blotting. f The K2A-1 cells were treated without or with daminozide (2.5 μM) for 24 h and the protein level of KDM2A, FAP and PDGFR-α was compared. g The association between KDM2A and FAP expression in the tumour stroma of 34 breast tumours, obtained from the microarray data GSE145148, was analysed and a positive correlation was found (P = 0.0341).