Ki 67

Parthenolide was purchased from MedChemExpress (cat# HY-N0141) and dissolved in DMSO. The Annexin V-FITC/PI staining kit was was purchased from Thermofisher (cat#88-8005-72). The antibodies for western blot are as followings: primary antibodies against phospho-EGFR (cat#3777), total-EGFR (cat#4267), phospho-ERK (cat#4377), total-ERK (cat#4695), phospho-AKT(T473) (cat#4060), phospho-AKT(T308) (cat#13,038), AKT (cat#9272), cleaved-caspase-3 (cat#9661), PARP (cat#9532), Ki-67(cat#9027), HRP goat anti-rabbit (cat#7074) and-mouse secondary antibodies (cat#5127) and GAPDH (cat#5174) were purchased from CST Cell Signaling Technology.

Frozen sections of lung tissue and cells were rinsed with PBS and fixed with 4% paraformaldehyde solution for 20 min. Tissue sections and cells were blocked with bovine serum albumin and incubated with the primary antibodies at 4°C overnight (vWF: 1 : 200, Zhongshan Jinqiao Biotechnology Co., Ltd., China; AAT1 1 : 50, Sigma-Aldrich, USA; p-p50 1 : 50 and ICAM-1 1 : 20, Santa Cruz Biotechnology Company, USA; p50 1 : 100 and Ki-67 1 : 100, Cell Signaling Technology Company USA; F4/80 1 : 300, α-SMA 1 : 300, collagen 1 : 25 and MCP-1 1 : 100, Abcam Company, UK). On the second day, the secondary fluorescent antibodies were incubated in the dark. All sections were counterstained with nuclear 4,6-diamino-2-phenylindole (DAPI; Zhongshan Jinqiao Biotechnology Co., Ltd., China). Immunofluorescence imaging was performed using a confocal laser scanning microscope (Leica, Wetzlar, Germany) for observation and comparison.