Figure 6. Anti-CD70-IFN-γ immunocytokines exert RIP1-dependent necrotic activity on RCC cell lines.
(A) RenCa, Caki-1, 786-O, or HRC63 cells were co-treated with bortezomib (MTD) and, respectively, murine (RenCa) or human (Caki-1, 786-O, and HRC63) IFN-γ immunocytokines (‘Anti-CD70 fusion’, 50 ng/ml) in the presence or absence of 50 μM RIP1 kinase inhibitor Nec-1 for 72–84 h. The MTD of bortezomib for 786-0 and HRC63 cells was 4 ng/ml and 2 ng/ml, respectively. Cell viability was determined by Trypan Blue exclusion analysis. Error bars represent mean +/− S.D; n = 3. (B) RenCa, Caki-1, 786-O, or HRC63 cells pre-treated without (-Nec-1) or with (+Nec-1) for 1h, before co-treatment with IFN-γ immunocytokines and bortezomib as in (A), were photographed 72 h post-treatment.
