Fig. 2.

Interaction between NSCLC cells and HUVECs in MCTSs affects the compactness of tumor spheroids. a and b NSCLC, which is stained by DiO, and HUVECs stained by DiD were cultured for 3 days in 2D or 3D culture systems and then the images were obtained. c NCI-H460 or A549 cells were co-cultured with HUVECs in 2D and 3D culture systems at a ratio of 5:5 for 48 h. Lysates were analyzed by immunoblotting using indicated antibodies. d NCI-H460, A549, and SK-MES-1 cells were co-cultured with stromal cells (WI38 cells and HUVECs) at a ratio of 5:5 for 1 day and 3 days. All bright-field images of spheroids were obtained using the Operetta® High Content Screening System with a 10× objective. e To calculate the volume of spheroids, their long and short diameters were measured. Data are shown as mean ± SD from two independent experiments in triplicate. *P < 0.05, **P < 0.01 versus Control (NCI-H460, A549, or SK-MES-1)