Fig. 1. Treatment with sunitinib activates NF-κB and upregulates expression of IL-6, IL-8 and TNF-α in ccRCC cells.

a Sunitinib (Sun) induces degradation of IκBα and promotes nuclear translocation of Rel-A in 786-O and PNX0010 ccRCC cells. Cells were treated with 10 μM of sunitinib for 3 h. Cytoplasmic and nuclear extracts were subjected to western blot analysis using specific antibodies. b 786-O and PNX0010 cells were treated with 10 μM of sunitinib for 4 h. NF-κB (p65) DNA binding activity was examined using TransAM assay. Data are presented as the mean ± S.D. ***P < 0.0001 when compared with cells cultured in medium only (Med). c 786-O cells were treated as described above. NF-κB activity was examined by EMSA in nuclear extracts prepared from 786-O cells. Supershift analysis using p50 antibody was performed to confirm that the observed bands are specific for NF-κB. d Sunitinib augments expression of IL-6, IL-8 and TNF-α in ccRCC cells. 786-O cells were treated with 10 μM of sunitinib for 10 h. The levels of IL-6, IL-8 and TNF-α were evaluated by ELISA (enzyme-linked immunosorbent assay). Data are presented as the mean ± S.D. ***P < 0.0005 when compared with cells cultured in medium only