Figure 1. Kidney cancer cell lines form cancer spheres with stem cell-related features in serum-free defined medium (SFDM) and after anti-miR-17 transfection.

(A) ACHN and CAKI-1 cells were propagated in SFDM or transfected with anti-miR-17, leading to the formation of 3D RCC spheres that could be propagated by enzymatic dissociation. Cells kept in regular (10% FBS) medium did not form spheres. (B) Sphere formation assay compared the self-renewal capacity of the parental cell lines and their 3D sphere derivatives. RCC spheres showed significantly higher sphere forming ability. (C) Stem cell marker expression of the parental cells, RCC spheres established in SFDM and spheres established by miR-17 inhibition was quantified by RT-qPCR. (D) Mesenchymal marker expression of parental cells, RCC spheres established in SFDM and spheres established by miR-17 inhibition was quantified by RT-qPCR. VIM: vimentin, SFDM: serum-free defined medium