Ki 67

The liver tissues of 6 groups of mice were fixed and sliced, with a thickness of about 4 μm. After antigen recovery, the slice was incubated with the primary antibody Ki-67 (1:50, CatNo: K009725P, Solarbio) diluted in PBS containing 3% albumin bovine V (CatNo. R4903, Solarbio) at 4 °C and then incubated with the secondary antibody diluted in PBS at room temperature for 60 min. After nuclear staining with DAPI (CatNo: C1002, Beyotime), the slides were mounted with a fade-resistant mounting medium. LSM 510 confocal microscopes were used to analyze immunofluorescence staining sections.

SLC27A1 (Solarbio, China, dilution:1:200), PTBP1 (Solarbio,China, dilution:1:100), EIF5A (Solarbio, China, dilution:1:50), Ki67 (Solarbio, China, dilution:1:100), VEGF (Solarbio, China, dilution:1:100), and P53 (Solarbio, China, dilution:1:200) were evaluated by immunohistochemistry. The paraffin-embedded specimens were cut into 4 μm thick sections. According to the manufacturer’s instructions, an antibody immunohistochemical analysis was performed on all sections using Leica Bond-III's automatic, random, and continuous slide staining system (Leica Biosystems, Germany). Images were obtained using a whole-slide scanner (3DHISTECH Ltd, Budapest, Hungary) and the immunohistochemical expression was scored by two observers using the CaseViewer software. The criterion of immunohistochemistry was that there were obvious brown granules in the cytoplasm or nucleus. The H score (range 0–300) was obtained by multiplying the staining intensity (0–3) by the percentage of positive cells (0%–100%), and the expression of tumor cells was scored semi-quantitatively.