Hematoxylin eosin h e staining

Male BALB/c nude mice (aging from 4 to 6 weeks, 18–22 g) were provided by the HFK Biotechnology company. All animal experimentation was approved by the Institutional Animal Care and Use Committee of Guangzhou Medical University. The A549 cell suspension (1 × 10⁶ cells/200 μL/mouse) containing PBS (Gibco, Invitrogen, USA) and Matrigel was fully re-suspended and injected into the axilla of the mice. The tumor-bearing mice were arbitrarily allocated to three groups. The volume of the tumor was estimated at regular intervals. Drug administration was started when the tumor volume was about 100 mm³. Full ultrasonic dissolution of the jaceosidin was dissolved into a uniform suspension (1% DMSO + 5% CMC-Na + normal saline) and intraperitoneal injected into mice (200 μL/mouse). The body weight and the tumor volumes were recorded every day. After 7 days of continuous administration, the mice were dissected and the tumors were excised. The tumor was fixed in 4% paraformaldehyde (Biosharp, China) for routine sections preparation. Routine sections were utilized for hematoxylin/eosin (HE) staining (Servicebio, China) and immunohistochemical (IHC) analysis according to the protocol [39 (link)]. The antibodies used in the IHC assay including E-cadherin (#3195, CST), ITGA2 (ab181548, Abcam), ITGB1 (ab179471, Abcam) and Vimentin (#5741, CST).