Ki67 antibody

Tissues were prepared for paraffin‐embedded sections. After that, the sections were dewaxed and rehydrated before treatment with 3% hydrogen peroxide solution. The sections were incubated with Ki67 antibody (1:50, Solarbio) or X‐ray repair cross‐complementing group 5 (XRCC5) antibody (1:100, Solarbio), followed by hatching with Goat Anti‐Rabbit IgG antibody (1:1000, GeneTex). After the sections were stained with hematoxylin, the Ki67 and XRCC5 positive cells were observed under a microscope.

Resected tumor tissues were fixed with 4% paraformaldehyde at 4°C for 24 h. The tissues were embedded in paraffin and sectioned (4 µm). After conventional dewaxing and rehydration with descending alcohol series at room temperature, the sections were incubated with 3% H₂O₂ for 15 min. The sections were incubated with 5% normal goat serum (SL038; Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 1 h. Subsequently, Ki-67 antibody (cat. no. GB121141; 1:1,000; Wuhan Servicebio Technology Co., Ltd.) or HMGCS1 antibody (cat. no. ab155787; 1:500; Abcam) were added for incubation at 4°C overnight after antigen retrieval with citric acid buffer (pH 6.0) for 1 min 40 sec at 100°C in a pressure cooker. Subsequently, the sections were incubated with HRP-conjugated secondary antibody (cat. no. G1214; 1:200; Wuhan Servicebio Technology Co., Ltd.) at room temperature for 1 h. The sections were rinsed three times using TBS with 0.1% Tween-20 for 5 min and visualized using DAB solution (cat. no. G1212; 1:1,000; Wuhan Servicebio Technology Co., Ltd.) for 10 min at room temperature. Images were acquired using a light microscope at a magnification of ×200.