Immunohistochemistry Analysis of Rab1A and FoxM1 in Colorectal Cancer

IHC was performed to investigate the expression of Rab1A and FoxM1 in 135 CRC tissue samples and adjacent normal tissues. All analyses were performed in accordance with the relevant guidelines and regulations. Surgical specimens were fixed in 10% formalin, embedded in paraffin, cut into 5-μm sections, dewaxed, and then rehydrated. Next, the processed sections were blocked with 10% goat serum for 30 min and incubated with the polyclonal antibodies of Rab1A and FOXM1 at room temperature overnight. IHC was performed using a tissue staining kit (Zhongshan Biotechnology, Beijing, China) following the manufacturer’s protocol. Immunostaining was independently examined by two clinical pathologists who were blinded to the patient outcome. The staining score was calculated using Yang, K’s method^{28 (link)}. The staining scores were as follows: 0 was deemed as (−),1–4 as (+), 5–8 as (++), and 9 –12 as (+++). In our current study, we classified all samples into a high expression group (++ or +++) or a low expression group (− or +) based on protein expression.