A549 cell line

The A549 cell line (Bioresource Collection and Research Center (BCRC) 60074) was purchased from the BCRC and cultured according to BCRC recommendations. For the cell viability and transfection assay, 4–5 × 10³ cells were seeded into the wells of a 96-well plate and cultured overnight. Cells were then treated according to the indicated doses and times. Cell viability was quantified using the CellTiter-Glo® Luminescent Cell Viability Assay kit (ATP assay) (Madison, WI, USA) as previously described [11 (link)]. The luminescent signal was measured using a photometer (SpectraMax® i3, Molecular Devices, Wals, Austria).

The Bm7 and Brmx2 human lung cancer cell line was established as previously described^{14 (link)}. The mouse lung cancer cell line TC-1 was kindly provided by Dr. T.C. Wu^{46 (link)}. A549 cell line was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan). Human umbilical vein endothelial cells (HUVECs) was kindly provided by Dr. Jeng-Jiann Chiu in National Health Research Institute, Taiwan. Human brain microvascular endothelial cells (HBMECs) and primary human astrocytes are purchased from ScienCell Research Laboratory (Carlsbad, CA, USA). Experiments were performed within 6 months of receipt from the cell bank to ensure cells’ ability. All cells used in this study were confirmed to be mycoplasma-free. The antibodies used in the study were against ADAM9 (MAB939, R&D Systems, Minneapolis, MN, USA), mouse ADAM9 (AF949, R&D Systems), ANGPT2 (AF623, R&D Systems), VEGFA (ABS82, Millipore, Billerica, MA, USA), CD31 (ab28364, Abcam), and elongation factor 1 α (EF1α, #05–235, Millipore). The activity of PLAT in different dilutions of the conditioned medium was detected with the AssaySense Human PLAT Immuno-Chromogenic Activity Assay Kit (Assaypro, St. Charles, MO) in 2-day cultured cells.