Fig. 6.

RCT001 inhibits proliferation of human RCC tumor spheres and synergize with anti PD-1. MEXF486 RCC tumors were grown as subcutaneous tumor xenografts in immunodeficient mice. After removal, the tumors are placed in layers of Versagel® from Cypre to achieve spheroid formation with human dermal fibroblasts (HDF). After the 3D tumors are established (7 days), they are treated with RCT001 (0.3 to 30 µM) for 7 days. a The total area of tumor spheroids was determined. b The number of tumor spheroids was determined. c—d Cell death was analyzed by DRAQ7 labelling. Quantification (C) and representative images (D) were shown. e–g Activated human PBMCs are first added to the cell culture medium (1 day before treatment); over time, they gradually infiltrate and attack the tumor spheres in the gel. Then tumor spheres + PBMCs were treated with RCT001 (3.3 and 10 µM) ± anti- PD -1 (pembrolizumab, 100 µg/ml). e The total area of tumorspheres was determined. f The number of tumorspheres was determined. g Cell death was analyzed by DRAQ7 labelling