Fig. 7

The IL-1β/IL1R1 system is involved in the invasive and proliferative features induced by hypoxia in MDA-MB-231 cells. a-b Cell spreading was evaluated in MDA-MB-231 cells transfected with scramble or AsGPER and exposed to normoxia or hypoxia (2% O2) in the presence or absence of IL1R1a. Cells were then trypsinized, plated onto coverslips coated with fibronectin and the morphology was recorded after 15 min (a) or 60 min (b). c-d Cell spreading was evaluated in MDA-MB-231 cells treated with vehicle and IL-1β alone or in combination with IL1R1a, as indicated. Cells were then trypsinized, plated onto coverslips coated with fibronectin and the morphology was recorded after 15 min (c) or 60 min (d). Scale bar 50 μM. Side panels show quantification of cell spreading, as indicated. Data are representative of three independent experiments performed in triplicate. e-f Transwell Matrigel invasion assay in MDA-MB-231 cells transfected with scramble or AsGPER and then cultured in normoxia or hypoxia (e) or treated with vehicle or IL-1β (f) alone or in combination with IL1R1a, as indicated. Cells were counted in at least 10 random fields in three independent experiments performed in triplicate, as quantified in the side panels. Scale bar 200 μM. g Spheroid formation assay in MDA-MB-231 cells exposed to hypoxia and IL-1β alone or in combination with IL1R1a. Scale bar 100 μm. Side panel shows the quantification of cell growth. Values represent the mean ± SD of three independent experiments performed in triplicate. (*), (○), (□) p < 0.05 for cells cultured under normoxia versus cells cultured under hypoxia or treated with IL-1β versus vehicle-treated cells