Fig. 1

Dihydroxylated bile acids CDCA and DCA destabilise HIF-1α in A-549, MCF-7 and DU-145 cells. BAs destabilise HIF-1α in (a) A-549 cells, (b) MCF-7 cells and (c) DU-145 cells. For fluorescent microscopy, all cells were grown under hypoxic conditions (DMOG 200 μM), except for untreated cells. All cells were stained for intracellular HIF-1α (green) using an anti-HIF1α antibody and 4′, 6-diamidino-2-phenylindole (DAPI) for DNA nuclear material (blue). HIF-1α staining was not observed in untreated cells whereas in DMOG treated cells, HIF-1α punctate staining was observed. Scale bar = 20 μM, magnification × 63 oil immersion. HIF-1α was quantitatively assayed in DU-145 cells using an anti-HIF-1α ELISA in the presence of BAs (d). Untreated cells exhibited reduced HIF-1α expression when compared to DMOG treated cells whereas in BA treated cells, HIF-1α levels were lower. e. Hexokinase II RT-PCR. Expression data normalised to HPRT is the average of two biological replicates (with two technical replicates) and is presented as fold change relative to the untreated control. Statistical analysis was performed using Repeated Measures ANOVA with post-hoc Bonferroni testing (* p < 0.05). For other data, statistical significance was assessed via an unpaired, two-tailed Student t tests. *, P < 0.05. Data presented represent those from three independent replicates