Fig. 1.

Secretomes from NSCLC spheroids induced endothelial-to-mesenchymal transition. a Experimental schematic of secretomes from NSCLC. NCI-H460 cells were cultured under 2D and 3D conditions using the same number of cells with the same amount of media. After 3 days, the conditioned medium (CM) was mixed with HUVEC original media at various ratios, and then used to treat HUVECs for 24 and 48 h. b Representative images of immunofluorescence staining for CD31 (green) and α-SMA (red) expression and nuclei (blue) in HUVECs treated with CM from 2D and 3D NCI-H460 cells at a ratio of 5:5. c HUVECs that were treated with CM from 2D and 3D NCI-H460 cells were harvested after 48 h incubation. HUVEC lysates were analyzed by western blotting using anti-CD31, anti-α-SMA, and anti-β-actin antibodies. d Quantification of relative expression levels of CD31 and α-SMA. Values were normalized to β-actin. Data are shown as mean ± SD from two independent experiments