Immunohistochemical Analysis of TYRO3 and ENO1

CRC and paired normal colorectal tissues were fixed using formalin, embedded in paraffin, cut into 5 μm sections, and stained using IHC [17 (link)]. The sections were incubated using the anti-TYRO3 (Abcam, UK) and anti-ENO1 (Boster, China) antibodies at 1:100 dilution for 2 hours at room temperature. TYRO3 expression in tissues was visualized with a tissue staining kit (Zhongshan Biotechnology, China). Two authors with expertise in pathology assessed the staining scores. IHC score was determined by the intensity multiple (0, negative; 1, weak; 2, moderate; 3, strong) and extent (0, 0–5%; 1, 6–25%; 2, 26–50%; 3, 51–75%; 4, >75%) score. We considered a final score of 0 as −; 1–4 as +; 5–8 as ++; 9–12 as +++. In this study, ++ or +++ was considered a positive expression, and – or + a negative [18 (link)].