Fig. 4

Knockdown of CXCL16 with siRNA caused activation of proliferation compared to the negative scrambled control in NSCLC cell lines A549 and NCI-H460. Cells were trypsinized briefly until detached, resuspended in complete growth media and counted. According to the manufacturer’s instructions, cells were seeded in duplicates into E-plates 16 after baseline measurement. Plates were incubated for 1 hour at room temperature, and then placed into the RTCA DP instrument located in an incubator preserving same temperature and CO₂ concentration as were used for routine cultivation of the cells. siRNA transfection mix was added to the cells 6 hours after seeding, and left there for 4 hours. Subsequently, the transfection mix was replaced with regular growth media. Cell index (arbitrary unit reflecting the cell-sensor impedance) was measured every 15 minutes during the first 4 hours for better resolution at attachment and spreading phase. Further measurements were taken every 30 minutes. Doubling times were calculated with RTCA software 1.2 (Roche). ± S.D of 4 technical replicates are shown. * P <0.001