Fig. 4.

Hypoxia-inducible factor (Hif)1a stabilization under Egln3 degradation promotes Hmox1 transcription

A. The top 20 DEGs between the normoxia and hypoxia groups and GSEA of the top 10 upregulated and downregulated signalling pathways. B. The protein levels of Egln3, Hif1a, and Hmox1 in MDFCs under normoxia and hypoxia. C. Western blot analysis of Egln3, Hif1a, and Hmox1 expression in MDFCs transduced with siEgln3. D. Representative fluorescence images showing the iron content and lipid peroxidation levels in MDFCs. Scale bar, 50 μm. E. 293T cells were transfected with 2000-, 1500-, 1300-, 1000- or 100-bp Hmox1 promoter-luciferase plasmids and further transfected with pcDNA3.1 or pcDNA3.1_HIF1A before the relative changes in luciferase activity were assessed. The binding site was mutated from ACGTGA to ATTTTA, and 293T cells were transfected with the mutant promoter and subsequently transfected with pcDNA3.1_HIF1A, followed by assessment of luciferase activity. F. ChIP assay to assess the binding of Hif1a to the Hmox1 promoter in MDFCs under normoxia and hypoxia. N.D., not detected. MDFCs, macrophage-derived foam cells; DEGs, differentially expressed genes; ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001.