Figure 5

Effect of periostin receptor silencing on NSCLC cells. (a,b) ITB3 knockdown A549 (a) or NCI-H520 (shITB3#1 and shITB#2) (b) cells and Mock-A549 or Mock-NCI-H520 cells were cultured for 6 days in 3D Matrigel containing DMEM with or without rh-periostin (200 ng/ml) and the area of the sphere was measured and evaluated. Upper panels show representative images of cell spheres and lower panels show results for sphere size. (c,d) The shITB3-A549 and shITB3-NCI-H520 (#1 and #2) cells were cultured for 6 days in 3D Matrigel containing DMEM or iCM with or without OC-20 (2 µg/ml), and the area of the sphere was measured. Mock-A549 or Mock-NCI-H520 cells were used as control. Statistical significance was tested with the Mann–Whitney U test. Solid squares show enlarged images and scale bars, 100 μm in (a–d). *P < 0.05, **P < 0.01. n.s., non-significant; ITB3, integrin β3; rh-periostin, recombinant periostin; iCM, diseased human lung fibroblast-IPF-derived conditioned media.