LXY6006 inhibits HIF-1α nuclear accumulation.

<p>(A) Nuclear extracts from T47D cells were subjected to Western blotting for HIF-1α and HIF-1β expression. HIF-1β was constitutively expressed and thus used as a loading control. (B) T47D cells pre-treated with or without 0.1 µM of LXY6006 were incubated in hypoxia conditions for different time, and then subjected to Western blotting. (C) T47D cells pretreated with or without 0.1 µM of manassantin A or LXY6006 were exposed to hypoxia (1%O₂, 8 hr) or 1,10-phenanthroline (phen) (10 µM, 8 hr), and then lysed for detection of nuclear HIF-1α expression as in (A). (D) MDA-MB-231, MX-1 and MX-1/Taxol cells were treated with or without 0.1 µM of manassantin A or LXY6006 and lysed for HIF-1α expression as above. (E) T47D cells treated with or without 0.1 µM of LXY6006 or manassantin A were exporesed to hypoxia for 8 hr, and then fixed for immunofluorescence straining for HIF-1α expression.