All GC tissue specimens were fixed with 10% neutral formaldehyde solution for 4–6 h at room temperature, and were routinely dehydrated, waxed and wrapped. Then, the tissues were cut into 3-µm sections. The tissue sections were baked at 60°C overnight. Then, the tissue sections were subsequently deparaffinized by the following method: After baking, the tissue slices were immersed in xylene at room temperature twice for 10 min each. Then they were immersed in 100% ethanol for 5 min. Next, the tissue sections were rehydrated in 100, 85 and 70% ethanol at room temperature for 2 min respectively. Finally, the slices were incubated at 90°C in deionized water for 30 min. Subsequently, the slices were boiled in citric acid buffer solution and maintained at a low heat for 30 min. The sections were then sealed with 5% BSA (Sangon Biotech Co., Ltd.) in a wet box at 37°C for 30 min and air-dried. The sections were then incubated in a wet box at 4°C overnight with an anti-RORβ antibody (cat. no. NBP1-82532; 1:300; Novus Biologicals, LLC). Following the primary antibody incubation, the sections were rinsed thrice with PBS for 5 min and incubated with 50 µl of an HRP-conjugated secondary goat anti-rabbit antibody (product code ab6721; 1:1,000; Epitomics; Abcam) at room temperature for 20 min. Then the sections were rinsed thrice with PBS for 3 min. The slides were observed under a light microscope with a magnification of ×100.

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