Ab270264

The rat prostate slices were de-waxed in xylene prior to ethanol gradient dehydration. Sections were received 10mM citrate buffer (pH, 6.0) for antigen retrieval and immersed in 3% hydrogen peroxide to block the endogenous peroxidase before immunohistochemistry (IHC). Sections were incubated with the primary antibodies against Ki-67 (2 µg/mL, ab15580, Abcam, Cambridge, MA, USA), IL-6 (1:400, ab6672, Abcam), IL-18 (1:2,000, ab223293, Abcam), and TNF-α (1:200, ab270264, Abcam) overnight at 4°C. Sections were washed with phosphatebuffered saline (PBS) thrice before and after incubation with the secondary antibody. After that, 1–3 minutes of diaminobenzidine color development was terminated. The nucleus of cells was stained by hematoxylin for 3 minutes, and sections were dehydrated, permeabilized, and sealed. Images were acquired using a light microscope (Nikon Eclipse E100, Tokyo, Japan).

The study was reviewed and approved by the Ethics Committee of The Second Affiliated Hospital of Wannan Medical College (Wuhu, China) and the ethical number is WYEFYLS2023138. The study was conducted in compliance with the principles of the Declaration of Helsinki (as revised in 2013). All participants provided full written informed consent.
The case inclusion criteria included age 18 years or above; newly diagnosed primary gastric tumor and Li confirmed by histology; without other treatments. The exclusion criteria included history of other malignancies. Three patients with newly diagnosed primary gastric tumor and Li who attended The Second Affiliated Hospital of Wannan Medical College were invited to participate in this study. All patients provided full written informed consent. We obtained three pairs of fresh tissue samples which contained primary gastric tumor tissues and Li tissues from these patients.
IHC was performed as previously described (26 (link)) on clinical specimens of gastric orthotopic tumor and Li using the primary antibodies against APOD (1:100, ab108191, Abcam, Cambridge, UK), JUN (1:100, ab40766, Abcam), CXCL5 (1:100, ARG66029, Arigo, Hsinchu, Taiwan, China), TNFAIP3 (1:100, ab270264, Abcam), IL7R (1:100, AG2272, Beyotime, Shanghai, China) and CD94 (1:100, ab235441, Abcam).