In this assay, we immunostained CA9, GLUT1, or GLUT3 in three different prostate/prostate cancer lines: immortalized RWPE1 cells, CTN-9 cells (a chemically transformed line derived from non-tumorigenic prostate epithelial PrEC-Hahn cells [52]), and DU145 (castrate-resistant brain metastatic tumor-derived prostate cancer line). Here, CA9 is linked to prostate cancer.