Figure 4
From: Anti-vascular agent Combretastatin A-4-P modulates Hypoxia Inducible Factor-1 and gene expression
CA-4-P induces NFkB nuclear translocation and activation. A: CA-4-P induces NFkB translocation from the cytoplasm to the nucleus. Western blot analysis of CA-4-P treated aerobic HUVEC demonstrating translocation of NFkB (p65) protein from the cytoplasmic to the nuclear fraction within 1 h in a dose dependent manner. C: cytoplasmic fraction, N: nuclear fraction. B: CA-4-P induces NFkB activation via Rho A, but not Rho kinase. EMSA showed that CA-4-P induced NFkB-DNA complex formation in aerobic HUVEC. Supershift analysis using antibodies against NFkB components resulted in a further retardation of the p50 band and loss of the p65 band. Treatment with Rho kinase inhibitors (Y27632 and HA1077) demonstrated no effect on NFkB, whereas treatment with the Rho A inhibitor C3 prevented NFkB activation by CA-4-P. (1) vehicle control, (2) 0.1mM CA-4-P, (3) + cold probe, (4) + p50 Ab, (5) + p65 Ab, (6) + Y27632, (7) + HA1077, (8) + C3.