Fig. 1.

Generation of primary cell cultures of RCC specimens. After surgical resection, the tumor tissue of clear cell RCC, papillary RCC and chromophobe RCC was minced, digested, filtered and cultivated. The cell cultures were validated using immunohistochemical staining for characteristic tissue- and tumor markers. For ccRCC specimens and their corresponding cultures, the VHL gene was sequenced, matching the VHL status of the cell cultures with the tissue they derived from. RNA was extracted from tumor and benign tissue for RNA-sequencing to identify central gene expression characteristics and alterations. All RCC subtypes were treated in vitro with tyrosine kinase inhibitors −/+2-deoxy-d-glucose (2-DG), using a CellTiter-Glo^® Luminescence assay and a crystal violet assay to assess the decrease of glycolysis and the cell viability after treatment; ccRCC clear cell renal cell carcinoma, VHLvon Hippel–Lindau gene