Fig 2: Expression of MB, CAIX, and LDHA in multiple organ tumor. Tissue microarrays containing cores obtained from multiple organ tumors and cancer adjacent normal tissues were stained with antibodies to MB, CAIX, and LDHA. All photomicrographs were obtained at 20× magnification and the scale bar indicates 50 µM. Black arrows indicate representative positive staining. (A) Squamous cell carcinoma (uterine cervix) with focal weak cytoplasmic MB expression, mild cytoplasmic and membranous CAIX expression, and strong diffuse cytoplasmic LDHA expression. (B) Duct adenocarcinoma (pancreas) with focal weak cytoplasmic MB expression in the acini, negative CAIX expression in the acini, and diffuse strong cytoplasmic and patchy membranous LDHA expression. (C) Adenocarcinoma of lung with patchy weak cytoplasmic MB expression in the acini, negative CAIX expression in the acini, and diffuse moderate to strong and patchy membranous and cytoplasmic LDHA expression. (D) Papillary carcinoma of thyroid with diffuse mild to moderate cytoplasmic MB expression in the acini, negative CAIX expression in the acini, and diffuse strong cytoplasmic LDHA expression.

Fig 3: Expression of MB, CAIX, and LDHA in glioblastoma multiforme. Tissue microarrays containing cores obtained from glioblastoma tissues, cancer adjacent normal cerebrum tissues, and normal cerebrum tissue were stained with antibodies to MB, CAIX, and LDHA. All photomicrographs were obtained at 20× magnification and the scale bar indicates 50 µM. Black arrows indicate representative positive staining. (A) Glioblastoma Multiforme (cerebrum) with focal weak cytoplasmic MB expression, negative CAIX expression, and patchy weak cytoplasmic LDHA expression. (B) Glioblastoma Multiforme (cerebrum) with moderate cytoplasmic MB expression, patchy strong cytoplasmic CAIX expression, and moderate cytoplasmic and occasional nuclear LDHA expression. (C) Glioblastoma Multiforme (cerebrum) with patchy weak cytoplasmic MB expression, patchy moderate cytoplasmic CAIX expression, and moderate diffuse strong cytoplasmic and occasional nuclear LDHA expression. (D) Glioblastoma Multiforme with diffuse mild to moderate cytoplasmic MB expression, focal moderate cytoplasmic CAIX expression, and diffuse moderate to strong cytoplasmic and few nuclear LDHA expression.

Fig 4: Carbonic Anhydrase 9 expression confirms the hypoxic condition. A-B: Representative images and densitometric analysis of Western blots for Carbonic Anhydrase (CA9) obtained from protein lysates of HCT116 and SW480 in normoxic conditions or subjected to hypoxic conditions. The graphs ordinate shows the OD (Optical Density) of the indicated proteins normalized for the housekeeping’s OD (ß-actin). Data are expressed as the mean ± SD of three independent experiments and statistical significance was analyzed using a Student’s t-test (* = p < 0.05; ** = p < 0.01). Corresponding uncropped full-length blots are included in Supplementary Materials

Fig 5: Expression of MB, CAIX, and LDHA in multiple organ tumor. Tissue microarrays containing cores obtained from multiple organ tumors and cancer adjacent normal tissues were stained with antibodies to MB, CAIX, and LDHA. All photomicrographs were obtained at 20× magnification and the scale bar indicates 50 μM. Black arrows indicate representative positive staining. (A) Cancer adjacent normal lung tissue with mild patchy cytoplasmic MB expression, negative CAIX, and focal weak cytoplasmic LDHA expression. (B) Cancer adjacent normal breast tissue with weak patchy cytoplasmic and focal nuclear MB expression, negative CAIX expression, and patchy moderate cytoplasmic, membranous and nuclear LDHA expression. (C) Cancer adjacent normal endometrial tissue with patchy weak to moderate cytoplasmic MB expression in the glands and occasional cytoplasmic in the endometrial stroma, negative CAIX expression in the glands and stroma, and diffuse moderate to strong cytoplasmic LDHA expression in the glands and mild to moderate cytoplasmic with patchy nuclear LDHA expression in the endometrial stroma. (D) Prostatic glands (hyperplasia) with focal mild cytoplasmic and membranous MB expression in the basal/myoepithelial cell layer only not in the luminal epithelial cells, negative CAIX expression in both basal and luminal cells, and patchy mild cytoplasmic and membranous LDHA expression in the basal/myoepithelial cell layer only not in the luminal epithelial cells.