Sc 35674

pCMV6-DDK-PDK1 and pCMV6-DDK (control vector) purchased from Origene (Rockville, MD) were transfected into OVCAR-3 using Lipofectamine 3000. After 72 h, transiently transfected cells were counted and plated for subsequent assays. For overexpression, stable clones were selected using G418 (500 µg/ml). OVCAR-3 cells stably overexpressing PDK1 were plated and transfected with siRNAs specific for α5 integrin (sc-29372), β1 integrin (sc-35674), or control (sc-37007) (Santa Cruz) for 48 h before cell counting, labeling, and plating for functional assays^{49 (link),50 (link)}.

ANGPTL2 (ab199133) antibody was purchased from Abcam (Cambridge, MA, USA), VEGF-A (A17877) was purchased from Abclonal (Cambridge, MA, USA), p-p65 (3033) antibody was purchased from Cell Signaling Technology (Danvers, MA, USA), β-actin (GT5512) and p65 (GTX102090) antibodies were purchased from GeneTex (Hsinchu, Taiwan). Small interfering RNAs (siRNAs) against integrin α5 (sc-29372) and β1(sc-35674), and immunoglobulin (Ig)-like transcript 4 (ILT4) (sc-45200) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). VEGF-A recombinant protein was acquired from PeproTech (Rocky Hill, NJ, USA). The p38 MAPK inhibitor SB203580 (HY-10256) and NF-κB inhibitor PDTC (P8765-1G) was acquired from Sigma-Aldrich (St. Louis, MO, USA). The ANGPTL2 overexpression plasmid (ANGPTL2 cDNA) was commercially synthesized by NCFB (Academia Sinica, Taiwan). ANGPTL2 (clone ID: TRCN0000158500) knockdown (sh-ANGPTL2) was purchased from the National RNAi Core Facility (RNAi Core, Academia Sinica, Taiwan).