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. Author manuscript; available in PMC: 2020 May 15.
Published in final edited form as: Cancer Res. 2019 Mar 21;79(10):2564–2579. doi: 10.1158/0008-5472.CAN-18-2674

Figure 4. Inhibiting EGLN1 increases HIF1A expression and reduces proliferation in EGLN1-dependent cells.

Figure 4.

A. Immunoblot showing that pharmaceutical inhibition(FG-4592) of EGLN1 increases HIF1A levels in a dose-dependent manner.

B. EGLN1 inhibitor increases HIF1A activity measured by using a Hypoxia Response Element (HRE) fused to a luciferase reporter. Luciferase activity was measured 48 hours post treatment with increasing dose of FG-4592. Data is representative of two independent experiments.

C. EGLN1 inhibitor reduces viability selectively in EGLN1-dependent cells. Cells were treated for 48 hours with increasing dose of FG-4592 (X-axis). Cell viability (Y-axis) was measured using CellTiter-Glo. The data shown is representative of three independent experiments.

D. EGLN1 inhibitor reduces long term proliferation in EGLN1-dependent cells. Results are representative of 3 independent experiments and data is graphed as mean ± standard deviation of 3 replicates.

E. EGLN1 inhibitor does not affect long term proliferation of EGLN1-insensitive cells. Results are representative of 3 independent experiments and data is graphed as mean ± standard deviation of 3 replicates.

F. EGLN1 inhibition reduces colony formation in a dose-dependent manner in EGLN1-dependent cells. Colonies were quantified as absorbance (Y-axis) over drug concentration (X-axis) over 10 days of treatment normalized to DMSO. Results are representative of 3 independent experiments.

G. EGLN1 inhibition has no significant effect on colony formation in EGLN1-insensitive cells. Colonies were quantified as absorbance (Y-axis) over drug concentration (X-axis) over 10 days of treatment normalized to DMSO. Results are representative of three independent experiments.

H. VHL inhibition reduces proliferation of EGLN1-dependent cells. Results are representative of 3 independent experiments, and data is graphed as mean ± standard deviation of 3 replicates. *p<0.05

I. VHL inhibition does not affect proliferation of EGLN1-insensitive cells. Results are representative of 2 independent experiments, and data is graphed as mean ± standard deviation of biological triplicates.