Fig 2: Circular diagram representation of PHD2-HIF1α interaction sites. Full-length UniProt sequences of PHD2 (Q9GZT9) and HIF1α (Q16665) were imported into the xiVew web portal with annotations of domain and regions of interests.

Fig 3: Analytical strategy for crosslinking-mediated identification of PHD2-HIF1α interaction sites. (A) A schematic representation of HTBH-tagged PHD2 for stable expression in HeLa cells. (B) Western blotting validation of HeLa cell lines with or without stable expression of HTBH-tagged PHD2 using PHD2 antibody and streptavidin (SA)-HRP. (C) Analytical workflow to identify in vivo HIF1α-PHD2 crosslinking sites with DSSO crosslinking and DMOG trapping strategies. Crosslinked peptides were identified through MS3 analysis triggered by signature fragmentations in MS2 spectra.

Fig 4: Quantitative profiling PHD2 interactome. (A) Venn diagram representing the overlap of PHD2 interacting proteins identified with and without DMOG treatment with label-free quantitative analysis. (B) Annotation enrichment analysis of Gene Ontology biological processes with FDR cutoff of 0.05. (C) Volcano plot of the label-free quantitative comparison of PHD2 interacting proteins with and without DMOG treatment (FDR<0.05) with selected genes highlighted in red (upregulation with DMOG treatment) and in blue (downregulation with DMOG treatment). (D) The interacting protein network of PHD2 interacting proteins that were upregulated after DMOG treatment. The interactions were extracted from STRING database through web portal.

Fig 5: EGLN1 inhibits the response to irradiation in CNE2 both in vitro and in vivo. A,B: To analyze the effect of EGLN1 on the survival of NPC cells after exposure to 6 single-radiation doses (0, 2, 4, 6, 8, and 10 Gy), their clonogenic ability was assessed. Quadratic and linear model equations were proposed for the analysis of radioresistance. C,D: Cell apoptosis in different groups was detected by flow cytometry analyses. Apoptosis rates decreased in EGLN1-OE groups and increased in ShEGLN1 groups compared with control. E, F: γH2AX was stained red, and the cell nucleus with DAPI (blue). The number of γH2AX was counted by analyzing 100 randomly selected cells. G: Representative tumor xenografts. H: Tumor volumes were measured every 3 d. I: Representative immunohistochemical images of caspase 3 detection and tunel assay in different groups are shown (×200). Data are presented as means ± SEM corresponding to three independent experiments. *P < 0.05; ***P < 0.005.