Vascular endothelial growth factor (vegf)

The antibodies used in the current study are listed in Table S1. The other reagents used are as follows: recombinant EGF, IL‐1β, IL‐7, IL‐13, IL‐15, GRO‐α, IL‐8, SDF‐1α, MCP‐3, MIP‐1α, MCP‐2, MIP‐3α, eotaxin‐3, bFGF, KGF, vEGF, IGF‐BP7, OPG, GM‐CSF (BBI Life Sciences, Shanghai, China); recombinant IL‐6, MMP‐3, TGF‐β1 (PeproTech); cetuximab (Merck KGaA); gefitinib, cisplatin, and LY3214996 (Selleck Chemicals Inc); MTT (3‐[4]‐2,5‐ diphenyltetrazolium bromide thiazolyl blue), X‐Gal (5‐bromo‐4‐chloro‐3‐indolyl‐β‐D‐galactopyranoside), DAPI (4′, 6‐diamidine‐2′‐phenylindole dihydrochloride), BrdU (5‐bromo‐2‐deoxyuridine), and PI (propidium iodide) (Sigma‐Aldrich); annexin V‐FITC/PI Apoptosis Detection Kit (Yeasen Biotech); Human IL‐8 Uncoated ELISA Kit (Thermo Scientific); Human Total MMP3 ELISA Kit and Human EGF ELISA Kit (Proteintech).

Cells were seeded in 96-well plates (10,000/well). After treatment with VEGF (BBI Life Sciences, Shanghai, China) for 24 h, cell viability was tested by cell counting kit-8 (CCK-8) (Dojindo, Kyushu, Japan). The viable cells were determined using standard enzyme instruments to detect the λ = 450 nm absorbance values.