AFfirm™ HIF1 alpha Mouse monoclonal Antibody - #BF8002

Fig. 2. ETV2 induces HIF-1α stabilization and nuclear accumulation by transcriptional inhibition of PHD2 and ERK1/2 phosphorylation (A) KEGG functional enrichment analysis of RNA sequence (B) A heatmap displays genes associated with osteogenesis and HIF-1 signaling (C, D) The protein and mRNA expression of HIF-1α and PHDs following ETV2 overexpression (E) Schematic of putative ETV2 binding elements on PHD2 promoter region (F) Dual luciferase reporter gene assay of ETV2 and PHD2 (G) Cytoplasmic HIF-1α, total and phosphorylated ERK1/2, and intracellular HIF-1α expression post ETV2 overexpression (H) Representative immunofluorescent images of intracellular phosphorylated ERK1/2. The Cy3 channel fluorescence represents lentiviral transfection. Scale bar: 50 μm (I) Representative immunofluorescent images of intranuclear HIF-1α. The Cy3 channel fluorescence represents lentiviral transfection. The orange arrows indicate the fluorescence of HIF-1α in the nucleus, while the white arrows point to the absence of HIF-1α fluorescence in the nucleus. Scale bar: 50 μm (NS, no significant difference, NC, negative control; OE, overexpression. Data are presented as the mean of >3 independent experiments ±SD. *P < 0.05, **P < 0.01, and ***P < 0.001).

Fig. 2. ETV2 induces HIF-1α stabilization and nuclear accumulation by transcriptional inhibition of PHD2 and ERK1/2 phosphorylation (A) KEGG functional enrichment analysis of RNA sequence (B) A heatmap displays genes associated with osteogenesis and HIF-1 signaling (C, D) The protein and mRNA expression of HIF-1α and PHDs following ETV2 overexpression (E) Schematic of putative ETV2 binding elements on PHD2 promoter region (F) Dual luciferase reporter gene assay of ETV2 and PHD2 (G) Cytoplasmic HIF-1α, total and phosphorylated ERK1/2, and intracellular HIF-1α expression post ETV2 overexpression (H) Representative immunofluorescent images of intracellular phosphorylated ERK1/2. The Cy3 channel fluorescence represents lentiviral transfection. Scale bar: 50 μm (I) Representative immunofluorescent images of intranuclear HIF-1α. The Cy3 channel fluorescence represents lentiviral transfection. The orange arrows indicate the fluorescence of HIF-1α in the nucleus, while the white arrows point to the absence of HIF-1α fluorescence in the nucleus. Scale bar: 50 μm (NS, no significant difference, NC, negative control; OE, overexpression. Data are presented as the mean of >3 independent experiments ±SD. *P < 0.05, **P < 0.01, and ***P < 0.001).

Figure 6 Nrf2 might protect bladder injury by activating its downstream antioxidant genes. (a) The protein expression of HO-1 in the bladder of the four groups. (b) The protein expression of NQO1 in the bladder of the four groups. Relative mRNA expression levels of HO-1 (c) and NQO1 (d) in the bladder of the four groups. ∗P < 0.05, ∗∗P < 0.01. Data are presented as mean ± SD.

Fig. 6. |TFP inhibits EMT of melanoma cells.D–F, IHC analysis of E-cadherin, vimentin, HIF1α, VEGFA, and Twist1 in melanoma tumor tissues of mice under stress stimulation. Representative images and staining scores are shown. Scale bar, 100 μm.

Fig. 4 |DRD2 overexpression inhibited the ubiquitin-dependent degradation of HIF1α. A, Venn diagram of proteins that interacted with DRD2 and HIF1α, as predicted by FPclass website. B, Molecular docking results of DRD2 and VHL. C, Co-IP results of DRD2 and HIF1α.

Fig. 5. The expression HIF-1α protein was enhanced in macrophages of human and mouse atherosclerotic lesions. (A): Dual immunofluorescence staining for HIF-1α (red), CD68 (green) and DAPI (blue) indicated the co-expression of HIF-1α and CD68 (a macrophage marker) in human normal artery free of AS and atherosclerotic lesions from LE and CA with or without DM. (B-C): Compared with histologically normal artery, the protein levels of HIF-1α were upregulated in human atherosclerotic lesions from LE and CA with or without DM by western blot analysis. (D): Dual immunofluorescence staining for HIF-1α (red), CD68 (green) and DAPI (blue) in ApoE-/- mice fed with a HFD or normal diet demonstrated that the HIF-1α expression was present in macrophages, as indicated by HIF-1α immunoreactivity colocalized with CD68. (E-F): Compared with ApoE-/- control mice, the protein levels of HIF-1α were upregulated in HFD-fed ApoE-/- mice by western blot analysis. *P