Fig. 4.

Relationships between the spatial heterogeneity of ⁶⁴Cu uptake and hypoxia and expression of copper transporters. a Three areas with different intensities for ⁶⁴Cu were distinguishable on the autoradiography: highly labeled area R1 (⁶⁴Cu++), an unstaining intermediate region R2 (⁶⁴Cu− and in the tumor periphery a moderate labeled region (⁶⁴Cu+). Autoradiography was co-registered with pimonidazole staining to evaluate severe hypoxia and also Hoechst 33342 nucleic acid stain to identify the tumor mass. Scale bar = 200 μm. b Representative immunostaining of hypoxia markers (HIF-1α, HIF-2α, CAIX, MCT-4), specific or non-specific copper transporters (CTR1, DMT1, and MDR1), redox state (SOD1), and different cell types (CD68 for monocytes/macrophages and GFAP for reactive astrocytes). The autoradiography and immunohistochemistry studies were performed on same animals and consecutive sections. Scale bar = 200 μm