Hematoxylin and eosin h e staining kit

L-Arginine (Art. No. JA2774) was purchased from Bomei Biotechnology Co., Ltd. (Hefei, China). The mouse interleukin-6 (IL-6) enzyme-linked immunosorbent assay (ELISA) kit (Art. No. KE10007, sensitivity 3.8 pg/mL, antibody specific for mouse IL-6) and the mouse tumor necrosis factor-α (TNF-α) ELISA kit (Art. No. KE10002, sensitivity 1.0 pg/mL, antibody specific for mouse TNF-α) were from ProteinTech (Wuhan, China). FITC-labeled dextran (Art. No. R-FD-006, Mr 100,000) was purchased from Xi’an Ruixi Biological Technology Co., Ltd. (Xi’an, China). Mouse high mobility group protein 1 (HMGB-1, Art. No. D721089-0096) ELISA kits were purchased from Sangon Biotech (Shanghai, China).The hematoxylin and eosin (H&E) staining kit (Art. No. ab245880) was purchased from Abcam (Cambridge, UK). The anti-HMGB-1 rabbit polyclonal anti-body (Art. No. D160488-0025, reactive to human, mouse and rabbit) was purchased from Sangon Biotech (Shanghai, China).The Alexa Fluor 488-labeled donkey anti-rabbit IgG (Art. No. A423) was purchased from Beyotime (Nanjing, China). Lipase activity assay kits (Art. No. A054-2-1) and α-amylase activity assay kits (Art. No. 100000060) were obtained from Jiancheng Biotech (Nanjing, China).

After metastasis examination by bioluminescent imaging assay, mice were euthanized and anatomized to obtain tissues from putative metastasis locations. Tissue samples were fixed by paraformaldehyde for 1 day and paraffin embedded. For osseous samples, tissues also underwent decalcification and dissection before paraffin treatment. Rotary Microtome (Leica, Frankfurt, Germany) was then utilized for tissue sample sectioning into pieces (5‐10 μm in thickness). After xylene‐deparaffinizing at 37°C for 20 min, sections were stained using Hematoxylin and eosin (H&E) Staining Kit (Sangon Biotech, Shanghai, China). BX53 Upright Microscope (Olympus, Tokyo, Japan) was used for morphological observation.