Figure 3.

LXY6090 inhibits HIF-1α protein accumulation by promoting its degradation.

Notes: MX-1 cells were treated with LXY6090 in the presence of 1 µM MG132 for 30 min and then cultured under hypoxic conditions for an additional 4 h. Whole cell extracts were prepared for Western blotting. The representative photographs and the quantification of HIF-1α/actin and VHL/actin levels normalized to the 21% O₂ group are shown (A). MX-1 cells were treated with/without LXY6090 at normoxic and hypoxic conditions; the hydroxyl-HIF-1α (Pro402 and Pro564), HIF-1α, and VHL proteins were determined by Western blotting. The representative photographs and the quantification of OH-HIF-1α/HIF-1α and VHL/actin levels normalized to the 21% O₂ group at each condition are shown. ***P<0.001 compared with the 21% O₂ non-treated group. ^#P<0.05, ^##P<0.01, ^### P<0.001 compared with the 1% O₂ non-treated group (B). VHL protein was knocked down by the shRNA method. Then, under hypoxic conditions, the effect of LXY6090 on VHL and HIF-1α was assessed by Western blotting. The representative photographs and the quantification of HIF-1α/actin and VHL/actin levels normalized to the non-treated group at each condition are shown. Data presented are of one representative experiment performed in triplicate. *P<0.05, **P<0.01 compared with the shRNA-control group (C).

Abbreviations: h, hours; HIF-1, hypoxia-inducible factor-1; min, minutes; VHL, von Hippel–Lindau.