Protein Expression of HIF1A and Ki67 in Lung Metastatic Colon Cancer

The expression of HIF1A and Ki67 at protein level in the experimental lung metastatic colon cancer tissues was detected by immunohistochemistry. Briefly, 5-μm-thick tissue sections were prepared. After deparaffinization, rehydration, antigen retrieval and blocking with H₂O₂ and 5% bovine serum albumin, respectively, the sections were incubated with primary antibodies against HIF1α (GeneTex Inc.) or Ki67 (Daco, Glostrup, Denmark) at 4°C overnight. After incubation with secondary antibody for 1 h at room temperature, DAB substrate (ZSGB Bio, Beijing, China) was used for staining. The staining score was given and calculated by allying intensity with extent as previously described (Wang et al., 2016b (link)).