Human gc tma

One human GC TMA was obtained from Outdo BioTech (Shanghai, China). In addition, we collected 10 GC samples from Zhongshan Hospital, Xiamen University (Xiamen, China) with approval by the medical ethics committee of Zhongshan Hospital, Xiamen University, and the corresponding written informed consent of all patients.
The detailed experimental procedures and immunohistochemical scores were described in our previous study [22 (link)]. For this study, we used anti-osteopontin (1:500 dilution; ab214050; Abcam, Cambridge, MA, USA), anti-CD68 (1:10,000 dilution; ab213363; Abcam), and anti-CD44 (1:400 dilution;3570; Cell Signaling Technology, Danvers, MA, USA) as the primary antibodies and calculated the immunohistochemical score based on the staining intensity score multiplied by the staining extent score.

Through Outdo BioTech, we acquired one human GC TMA (Shanghai, China). Moreover, our prior work described the specific experimental methods and immunohistochemistry scores used to evaluate the samples [3 (link)]. Briefly, gastric cancer and paracancerous tissues were fixed in 10% formalin, paraffin-embedded, sliced into 4-6 um sections, and placed onto slides. After deparaffinization, rehydration, and microwave antigen retrieval, the slides were incubated with primary antibodies against CD20 (1:5000 dilution; Proteintech Group, Inc., Rosemont, IL, USA), CD19 (1:5000 dilution; Proteintech Group, Inc., Rosemont, IL, USA), and CXCR4 (1:1000 dilution; ab214050; Abcam, Cambridge, MA, USA) antibodies at 4 °C overnight. Afterward, the slides were incubated with a secondary antibody at room temperature for 30 min and stained with DAB substrate, followed by hematoxylin counterstaining. The immunohistochemical score was determined by multiplying the staining intensity score by the staining extent score. The positive result is a brown color of varying shades. The color depth of brown was positively correlated with the protein expression level.