Figure 1

Mature adipocytes promote tumor invasion in a CFD dependent manner. (A) Schematic illustration of the transwell migration/invasion assays in which adipocytes and cancer cells were cultured in the lower well and upper chamber, respectively. Mature adipocytes were prepared by inducing the differentiation of confluent ADSCs for 8 days (Middle and right). Adipocyte precursors (ADSCs) were seeded the day before the analysis (Left). For invasion assays, upper chambers were prepared by coating the upper surface of the filter membrane with DMEM with 10% Matrigel, while the filter membrane remained uncoated for migration assays. Upper chambers seeded with breast cancer cells were put in the wells and migration/invasion abilities of breast cancer cells were evaluated. (B, C) Invasion ability of murine breast cancer EO771 cells cocultured with WT or Cfd KO murine ADSCs (B) or mature adipocytes (C). Left: representative images of the invaded cells. Black arrows: pore in the transwell membrane, white arrows: transmigrated cells. Scale bars, 50 μm. Data are presented as mean ± SD. n = 4, *p < 0.05. (D, E) Invasion ability of human breast cancer MDA-MB-231 cells cocultured with human ADSCs (D) or mature adipocytes (E) supplemented with or without 10 μM danicopan, a competitive inhibitor for CFD (CFD Inh). Left: representative images of the invaded cells. Black arrows: pore in the transwell membrane, white arrows: transmigrated cells. Scale bars, 50 μm. Data are presented as mean ± SD. n = 4, *p < 0.05.