Fig. 3

Lung tumour CM and secreted-proteins enhance adhesion of A549 and SK-MES-1 cells to human brain endothelial monolayers. Confluent hCMEC/D3 cells in μ-slides were exposed to A549 CM or SK-MES-1 CM for 30 min and 24 h prior to perfusion with calcein AM stained A549 (a) or SK-MES-1 cells (c). A549 and SK-MES-1 adhesion to hCMEC/D3 cells treated with 80 ng/ml CC, 10 ng/ml CL, 200 ng/ml IGFBP-7, 0.2 ng/ml VEGF and 160 pg/ml TNF-α at 4 and 24 h (b) and (d). Mean values from three separate experiments were statistically analysed. * p ≤ 0.05 versus control levels (100 %). e Representative images of adherent calcein-AM stained A549 [(i) and (ii)] and SK-MES-1 [(iii) and (iv)] cells to DMEM-BS [(i) and (iii)] and CL-treated [(ii) and (iv)] brain endothelial monolayers. Scale bar = 75 μm