Under hypoxia, HIF-1a is stabilized and moves into the nucleus, where it regulates transcription of genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia.
Figure 1. HIF-1a staining of HeLa cells. Anti-HIF-1a antibody [EPR16897] (ab179483) was used at 1/50 dilution with goat anti-rabbit IgG (Alexa Fluor® 488) secondary antibody. Cells were treated with 0.6 nM CoCl2 for 6 hours.
|Western blot, immunocytochemistry||Anti-HIF-1a antibody (ab179483)|
|ELISA||Human SimpleStep ELISA® kit (ab171577)|
|Positive control||Recombinant human HIF-1a protein (ab154478)|
|Activity assay||HIF-1a transcription factor assay kit (ab133104)|
BNIP3 induction by HIF-1α under hypoxic conditions is involved in hypoxia-mediated autophagy, cell survival and tumor progression1.
Figure 2. BNIP3 staining of laryngeal squamous cell carcinoma tissue. Anti-BNIP3 mouse monoclonal antibody (ab10433) was used at 1/100 dilution and visualized by DAB.
|Various||Anti-BNIP antibody [ANa40] (ab10433)|
|ELISA||Human HIF-1a + BNIP3 hypoxia in cell ELISA kit (ab129733)|
|Flow cytometry||HIF-1a + BNIP3 hypoxia response human flow cytometry kit (ab126585)|
PDK1 (pyruvate dehydrogenase kinase 1) is downstream of HIF-1α and is induced under hypoxia. Expression during hypoxia channels glucose through glycolysis to maintain ATP production2.
Figure 3. PDK1 staining of NIH/3T3 cells. Anti-mitochondrial pyruvate dehydrogenase kinase 1 antibody (ab202468) was used at a dilution of 1/1000 with goat anti-rabbit IgG (Alexa Fluor 488) secondary antibody (ab150077). Cells were counterstained with DAPI (blue) and tubulin also stained (red).
|Western blot, ICC, IP, flow cytometry||Anti-mitochondrial pyruvate dehydrogenase kinase 1 antibody (ab202468)|
|ELISA||Human HIF-1a + PDK1 hypoxia in cell ELISA kit (ab125299)|
|Flow cytometry||HIF-1a + PDK1 hypoxia response human flow cytometry kit (ab126700)|
|Positive control||Recombinant human mitochondrial pyruvate dehydrogenase kinase 1 protein (ab125580)|
Expression of the GLUT1 glucose transporter is increased under hypoxic conditions, induced by HIF-1α3.
Figure 4. GLUT1 staining of HEPg2 cells. Anti-glucose transporter GLUT antibody (ab115730) was used at a dilution of 1/100 with Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150077). Cells were counterstained with DAPI and tubulin was also stained (red).
|Western blot, ICC/IHC, flow cytometry||Anti-glucose transporter GLUT antibody (ab115730)|
|ELISA||Human HIF-1a + GLUT hypoxia in cell ELISA kit (ab125298)|
|Flow cytometry||HIF-1a + GLUT1 hypoxia response human flow cytometry kit (ab126584)|
1. Bellot G, Garcia-Medina, Gounon P, Chiche J, Roux D, Pouysségur J, Mazure NM. Hypoxia-induced autophagy is mediated through hypoxia-inducible factor induction of BNIP3 and BNIP3L via their BH3 domains. Mol Cell Biol. 10, 2570–81 (2009).
2. Kim JW, Tchernyshyov I, Semenza GL, Dang CV. HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia. Cell Metab. 3, 177–85 (2006).
3. Chen C, Pore N, Behrooz A, Ismail-Beigi F, Maity A. Regulation of glut1 mRNA by hypoxia-inducible factor-1. Interaction between H-ras and hypoxia. J Biol Chem. 276, 9519–25 (2000).