Journal: Cancer Cell International

Article Title: ARNT is a potential direct HIF-1 target gene in human Hep3B hepatocellular carcinoma cells

doi: 10.1186/s12935-017-0446-2

Figure Lengend Snippet: Materials

Article Snippet: Anti-HIF-1α , Mouse monoclonal, clone 54, #610959, BD Transduction Laboratories™ , WB.

Techniques: Transduction

Journal: Cancer Cell International

Article Title: ARNT is a potential direct HIF-1 target gene in human Hep3B hepatocellular carcinoma cells

doi: 10.1186/s12935-017-0446-2

Figure Lengend Snippet: Recruitment of HIF-1α and ARNT to the ARNT gene promoter. a Functional elements within the ARNT promoter sequence upstream to −1200 bp from the transcription start site (bp 1). HBS HIF binding site, HAS HIF ancillary sequence, TATA TATA box. Investigated partial sequences are indicated (Region 1, Region 2, Region 3). b Chromatin immunoprecipitation (ChIP) assays. Values are presented as mean ± SEM of n = 3 independent experiments. IgG normal rabbit IgG, H3 histone H3

Article Snippet: Anti-HIF-1α , Mouse monoclonal, clone 54, #610959, BD Transduction Laboratories™ , WB.

Techniques: Functional Assay, Sequencing, Binding Assay, Chromatin Immunoprecipitation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Association of Age with the Expression of Hypoxia-Inducible Factors HIF-1α, HIF-2α, HIF-3α and VEGF in Lung and Heart of Tibetan Sheep

doi: 10.3390/ani9090673

Figure Lengend Snippet: Immunostaining of four proteins in the heart of Tibetan sheep. All sections are from two-year-old sheep. ( A , B ): Hypoxia-inducible factor (HIF)-1α immunostaining in the heart; ( C , D ): HIF-2α immunostaining in the heart; ( E , F ): HIF-3α immunostaining in the heart; ( G , H ): Vascular endothelial growth factor (VEGF) immunostaining in the heart; ( I – L ): control of HIF-1α, HIF-2α, HIF-3α and VEGF in heart respectively. Arrows in ( B , D , H ): HIF-1α, HIF-2α and VEGF immunostaining in myocardial nuclei, respectively. The positive immunostaining showed brown with non-immunostained nuclei blue. ( A , C , E , G , I ): 20×; ( B , D , F , H , J – L ): 100×).

Article Snippet: Subsequently, they were incubated in a humid chamber overnight at 4 °C with a primary antibody of rabbit anti-HIF-1α, anti-HIF-2α, anti-HIF-3α or anti-VEGF polyclonal antibody (all antibodies diluted 1:200, Bioss, China, bs-20398R, bs-1447R, bs-5989R and bs-1665R), as described previously [ ].

Techniques: Immunostaining

Journal: Animals : an Open Access Journal from MDPI

Article Title: Association of Age with the Expression of Hypoxia-Inducible Factors HIF-1α, HIF-2α, HIF-3α and VEGF in Lung and Heart of Tibetan Sheep

doi: 10.3390/ani9090673

Figure Lengend Snippet: Immunostaining of four proteins in the lung of two-year-old Tibetan sheep. ( A – C ): Hypoxia-inducible factor (HIF)-1α immunostaining in the lung; ( D – F ): HIF-2α immunostaining in the lung; ( G–I ): HIF-3α immunostaining in the lung; ( J–L ): Vascular endothelial growth factor (VEGF) immunostaining in the lung; ( M– P ): control of HIF-1α, HIF-2α, HIF-3α and VEGF in lung, respectively. Arrows in ( C , F , I , L ): HIF-1α, HIF-2α, HIF-3α and VEGF immunostaining in alveolar macrophages, respectively. The positive immunostaining was brown with non-immunostaining nuclei blue. ( A , D , G , J , M , O , P ): 20×; ( B , C , E , F , H , I , K , L , N ): 100×).

Article Snippet: Subsequently, they were incubated in a humid chamber overnight at 4 °C with a primary antibody of rabbit anti-HIF-1α, anti-HIF-2α, anti-HIF-3α or anti-VEGF polyclonal antibody (all antibodies diluted 1:200, Bioss, China, bs-20398R, bs-1447R, bs-5989R and bs-1665R), as described previously [ ].

Techniques: Immunostaining

Journal: Animals : an Open Access Journal from MDPI

Article Title: Association of Age with the Expression of Hypoxia-Inducible Factors HIF-1α, HIF-2α, HIF-3α and VEGF in Lung and Heart of Tibetan Sheep

doi: 10.3390/ani9090673

Figure Lengend Snippet: The IOD (Integrated optical density) value of hearts and lungs of Tibetan sheep of different ages. VGEF is vascular endothelial growth factor, HIF is hypoxia-inducible factor. Values are means ± SD ( n = 10). * indicates that significant differences ( p < 0.05) in the IOD values existed between immunostaining intensity for VEGF and all three HIFs at that time point. + indicates significant differences in the immunostaining intensity of VEGF and HIF-1α in the heart and HIF-1α HIF-2α in the lung between 6-year-old and 1-year-old sheep.

Article Snippet: Subsequently, they were incubated in a humid chamber overnight at 4 °C with a primary antibody of rabbit anti-HIF-1α, anti-HIF-2α, anti-HIF-3α or anti-VEGF polyclonal antibody (all antibodies diluted 1:200, Bioss, China, bs-20398R, bs-1447R, bs-5989R and bs-1665R), as described previously [ ].

Techniques: Immunostaining

Journal: Animals : an Open Access Journal from MDPI

Article Title: Association of Age with the Expression of Hypoxia-Inducible Factors HIF-1α, HIF-2α, HIF-3α and VEGF in Lung and Heart of Tibetan Sheep

doi: 10.3390/ani9090673

Figure Lengend Snippet: Hypoxia-inducible factor (HIF)-1α, HIF-2α, HIF-3α and vascular endothelial growth factor (VEGF) expression in the heart and lung of Tibetan sheep were measured by ELISA. Values are means ± SD ( n = 10). * indicates a significant difference in expression between 1-year-old and 6-year-old sheep ( p < 0.05).

Article Snippet: Subsequently, they were incubated in a humid chamber overnight at 4 °C with a primary antibody of rabbit anti-HIF-1α, anti-HIF-2α, anti-HIF-3α or anti-VEGF polyclonal antibody (all antibodies diluted 1:200, Bioss, China, bs-20398R, bs-1447R, bs-5989R and bs-1665R), as described previously [ ].

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Animals : an Open Access Journal from MDPI

Article Title: Association of Age with the Expression of Hypoxia-Inducible Factors HIF-1α, HIF-2α, HIF-3α and VEGF in Lung and Heart of Tibetan Sheep

doi: 10.3390/ani9090673

Figure Lengend Snippet: Hypoxia-inducible factor (HIF)-1α, HIF-2α, HIF-3α and vascular endothelial growth factor (VEGF) gene expression in the heart and lung of Tibetan sheep were measured by RTPCR (Reverse transcription polymerase chain reaction). Values are means ± SD ( n = 10). * indicates a significant difference ( p < 0.05) in gene expression within a tissue between 1-year-old and 6-year-old sheep. + indicates a significant difference ( p < 0.05) between gene expression in the heart and the expression of the same gene at the same age in the lung. The bar containing + has the significantly higher gene expression.

Article Snippet: Subsequently, they were incubated in a humid chamber overnight at 4 °C with a primary antibody of rabbit anti-HIF-1α, anti-HIF-2α, anti-HIF-3α or anti-VEGF polyclonal antibody (all antibodies diluted 1:200, Bioss, China, bs-20398R, bs-1447R, bs-5989R and bs-1665R), as described previously [ ].

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Foods

Article Title: Sea Buckthorn Pericarp Flavonoids Improve Diet-Induced Hyperlipidemia via Coordinated Modulation of Hepatic Lipid Metabolism and Gut Microbiota

doi: 10.3390/foods15061049

Figure Lengend Snippet: TFSP modulates the expression of ACC, FAS, CPT-1α, PPARα and ATGL proteins in the livers of HFD mice. n = 6 per group. Data are presented as mean ± SD. Different letters above bars indicate statistically significant differences ( p < 0.05) by one-way ANOVA followed by LSD post hoc test.

Article Snippet: Peroxisome proliferator-activated receptor alpha (PPARα) primary antibody (Cat. No. A00600-2), carnitine palmitoyltransferase-1 alpha (CPT-1α) primary antibody (Cat. No. A00917-3), acetyl-CoA carboxylase (ACC) primary antibody (Cat. No. M01802-2), fatty acid synthase (FAS) primary antibody (Cat. No. BA0484), adipose triglyceride lipase (ATGL) primary antibody (Cat. No. A01800-1), and GAPDH primary antibody (Cat. No. BM1623) were obtained from BOSTER Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Oncogene

Article Title: BATF2 prevents glioblastoma multiforme progression by inhibiting recruitment of myeloid-derived suppressor cells

doi: 10.1038/s41388-020-01627-y

Figure Lengend Snippet: A ELISA detection of MDSCs-associated cytokines in U251-Ctrl and U251-BATF2 tissues ( n = 3, independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001). B ELISA detection of SDF-1α in 7, 14, and 21 days in U251-Ctrl and U251-BATF2 tumour ( n = 3, independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001). C Representative IHC images of BATF2 and SDF-1α on consecutive sections of U251-Ctrl and U251-BATF2 intracranial tumours. Scale bars, 100 μm. D Representative images of immunofluorescent staining with SDF-1α (green), SV40 large T (red) and DAPI (blue) in U251-Ctrl and U251-BATF2 intracranial tumours. Scale bars, 25 μm. E Percentages of CXCR4 and CXCR7 expressing cells in CD11b + Gr1 + MDSCs isolated from tumours by FACs analysis. F Representative image of U251 tumour injected with U251-Ctrl- and U251-BATF2-derived EVs for four times, the subcutaneous tumour volume and SDF-1α detection by ELISA ( n = 3 independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001). G Western blot detection of HIF-1α and SDF-1α in 7, 14 and 21 days during U251 tumour progression. H EVs labelled by PKH67 added to phalloidin staining U251 cells and western blot detection of HIF-1α and SDF-1α. Scale bars, 20 μm. I ELISA and qPCR detection of SDF-1α in U251 tumour treated with Ctrl-EVs and BATF2-EVs ( n = 3, independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001). J ELISA detection of SDF-1α with and without 1% Triton X-100 under hypoxia condition ( n = 3, independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001). K Nano-FCM analysis and statistics of SDF-1α expression in Ctrl-EVs and BATF2-EVs ( n = 3, independent experiment, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Antibodies used in this study were as follows: BATF2 (ab157466, Abcam, London, UK); BATF2 (Thermo Fisher, PA5-43094); BATF2 (241887, Santa Cruz Biotechnology, CA, USA); CD63 (Abcam, ab217345); CD9 (Abcam, ab92726); TSG101 (Abcam, ab125011); SDF-1α (Abcam, ab9797); SDF-1α (Abcam, ab181018); HIF-1α (H6536, Sigma); CXCR4 (Abcam, ab135170); VEGF (ab46154, Abcam); CD33 (Abcam, ab203253); CD14 (Abcam, ab203294); Gr-1 (108448, Biolegend); CD11b (Abcam, ab13357); CXCR7 (Biolegend, #391405); SV40 Large T Antigen (CST, #15729, CA, USA); MMP-9 (CST, #13667); 10 nm gold-conjugated goat anti-rabbit secondary antibody (bs-0437P-Gold, Bioss, China), GAPDH (sc365062, Santa Cruz Biotechnology), and β-Actin (sc-47778, Santa Cruz Biotechnology).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Isolation, Injection, Derivative Assay, Western Blot

Journal: Oncogene

Article Title: BATF2 prevents glioblastoma multiforme progression by inhibiting recruitment of myeloid-derived suppressor cells

doi: 10.1038/s41388-020-01627-y

Figure Lengend Snippet: A Representative images of BATF2, CD33, CD14, SDF-1α, and HIF-1α in human malignant glioma tissues from 50 patients. Scale bars, 100 μm. B Correlations analyses of BATF2, CD33, CD14, SDF-1α, and HIF-1α in human malignant glioma tissues from 50 patients. The Pearson’s correlation coefficient ( r ) and p value are shown ( n = 50). C Exo-Counter detection of BATF2 + EVs in tumour tissues of I–II GBM, III–IV GBM and healthy donors ( n = 5, * p < 0.05, ** p < 0.01, *** p < 0.001). D Exo-Counter detection of BATF2 + EVs in plasma of I-II GBM, III-IV GBM and healthy donors ( n = 24, * p < 0.05, ** p < 0.01, *** p < 0.001). E ROC curve of BATF2 + EVs numbers data from I–II GBM vs. healthy donors, III–IV GBM vs. healthy donors, I–II GBM vs. III–IV GBM ( n = 24, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Antibodies used in this study were as follows: BATF2 (ab157466, Abcam, London, UK); BATF2 (Thermo Fisher, PA5-43094); BATF2 (241887, Santa Cruz Biotechnology, CA, USA); CD63 (Abcam, ab217345); CD9 (Abcam, ab92726); TSG101 (Abcam, ab125011); SDF-1α (Abcam, ab9797); SDF-1α (Abcam, ab181018); HIF-1α (H6536, Sigma); CXCR4 (Abcam, ab135170); VEGF (ab46154, Abcam); CD33 (Abcam, ab203253); CD14 (Abcam, ab203294); Gr-1 (108448, Biolegend); CD11b (Abcam, ab13357); CXCR7 (Biolegend, #391405); SV40 Large T Antigen (CST, #15729, CA, USA); MMP-9 (CST, #13667); 10 nm gold-conjugated goat anti-rabbit secondary antibody (bs-0437P-Gold, Bioss, China), GAPDH (sc365062, Santa Cruz Biotechnology), and β-Actin (sc-47778, Santa Cruz Biotechnology).

Techniques:

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: (A) In uninfected skin, rare and faint nuclear HIF-1α staining occurred in the epidermis whereas the dermis was negative for HIF-1α (A1). (B to L) In all cutaneous infections, a strong nuclear HIF-1α signal was detected in keratinocytes, predominantly of the spinal cell layer (examplified in L). Moreover, endothelial cells of dermal capillaries frequently stained positive (arrows in B1, C1, D1, E1, G, I1). Arrowheads: (B2) border zone of the positive abscess; (C2): positive neutrophils; (F): positive dermal lympho-histiocytic infiltrates; (F1): large positive nuclei of a multi-nuclear giant cell; (F2): skin lesion surrounded by a HIF-1 positive inflammatory infiltrate; (H): positive sub-corneal neutrophils (higher power in H1); (I): positive intra-epidermal infiltrate; (K): intradermal nodules containing leishmaniae. (A) Control: uninfected sample; (B–E): bioptic samples of patients suffering from infections with humanpathogenic bacteria [(B) S. aureus , (C) S. agalactiae and S. aureus , (D) A. baumanii and E. coli and (E) B. burgdorferi ], (F, G) with humanpathogenic Herpes-viruses [(F) Varicella zoster virus (VZV) and (G) Human Herpes Virus-8 (HHV 8)], (H–J) with humanpathogenic fungi [(H+I) T. rubrum and (J) C. albicans ] or (K, L) with the protozoic pathogen L. donovani . Magnifications: all 250×, except for A1, B1, B2, C1, D1, E1, F2, I1 (all 400×), and C2, F1, H1, L (all 1,000×).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Staining, Control, Bacteria, Virus

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: Four to six hours upon infection, HIF-1 activation was determined by Western blotting (loading control: actin). pO 2 levels were quantified in the medium of control and infected cells (A: S. aureus ATCC 33592, MOI 20, infection time: six hours; B: S. epidermidis ATCC 12228, MOI 20, infection time: six hours; C: E. coli ATCC 25992, MOI 10, infection time: four hours). Negative control: uninfected cells, positive control: hypoxia or DFO (200 µmol/L).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Infection, Activation Assay, Western Blot, Control, Negative Control, Positive Control

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: Four to six hours upon infection, HIF-1 activation was determined by Western blotting (loading control: actin). pO 2 levels were quantified in the medium of control and infected cells (D: P. aeruginosa ATCC 27853, MOI 10, infection time: four hours; E: S. agalactiae SK 43, MOI 200, infection time: five hours; F: C. albicans ATCC 90028, MOI 20, infection time: six hours). Negative control: uninfected cells, positive control: hypoxia or DFO (200 µmol/L).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Infection, Activation Assay, Western Blot, Control, Negative Control, Positive Control

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: Six hours upon infection, HIF-1 activation was analyzed in (A) Western blots (loading control: actin) and by determining (B) the induction of the HIF-1-dependent luciferase activity. HeLa-229 cells were transfected with a 4xHRE-luc reporter construct 24 hours before infection (triplicate means given; for details see ). Induction was determined by chemiluminescence. Bacterial induced (C) VEGF and (D) HIF-1α transcription. Total mRNA was prepared six hours upon infection, transcribed into cDNA and quantified by real-time Light-Cycler-PCR (ratio: VEGF/actin or HIF-1α/actin transcripts; triplicate means given). Negative control: uninfected cells; S. aureus wt (8325-4); S. aureus Δ hemB ( S. aureus ermB Ω hemB 8325-4, SCV); S. aureus Δ hemB , hemB -complemented ( S. aureus pCX19Ω hemB 8325-4, complemented SCV mutant) ; positive control: DFO (200 µmol/L). Multiplicity of infection (MOI): 20. * significant difference to control cells (P<0.05).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Infection, Activation Assay, Western Blot, Control, Luciferase, Activity Assay, Transfection, Construct, Negative Control, Mutagenesis, Positive Control

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: The HIF-1α protein was analyzed in Western blots six hours upon infection (loading control: actin) of HeLa 229 cells with the following bacterial strains: (A) S. aureus 8325-4 (wt), S. aureus ermB Ω hemB 8325-4 (SCV), S. aureus SA113 (wt), S. aureus SA113 Δ ypf::ermB (reduced in lipoteichoic acid synthesis), S. aureus SA113 Δ tagO (defective in producing wall teichoic acid); (B) S. aureus ATCC 25923 (wt), S. epidermidis ATCC 12228 (wt), S. aureus Newman (wt), S. aureus Newman mAH12 (defective in producing the extracellular adherence protein EAP). Negative control: uninfected cells, positive control: DFO (200 µM). Multiplicity of infection (MOI): 20.

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Western Blot, Infection, Control, Negative Control, Positive Control

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: The HIF-1α protein was analyzed in Western blots six hours upon infection (loading control: actin). The following patient isolates were used: S. aureus patient isolate 1 (wt; S. aureus A22223 I), S. aureus patient isolate 2 (clinical SCV; S. aureus A22223 II), S. aureus patient isolate 3 (wt; S. aureus OM 1a), S. aureus patient isolate 4 (clinical SCV; S. aureus OM 1b). Negative control: uninfected cells, positive control: hypoxia. Multiplicity of infection (MOI): 20.

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Western Blot, Infection, Control, Negative Control, Positive Control

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: (A) Control and infected HeLa-229 cells were incubated with the hypoxic marker pimonidazole hydrochloride which was visualized by using specific Cy3 labelled antibodies (for details see ). Scale bare: 40 µm. (B) pO 2 levels were quantified in the medium of control and infected cells (triplicate means given). (C) Hela-229 cells were incubated in gas permeable (gas-perm) and conventional (conv) culture dishes and HIF 1α protein was analyzed in Western blots (loading control: actin). (D) pO 2 levels in the medium of control or S. aureus infected HeLa-229 cells (means of doubles). Negative control: uninfected cells; S. aureus wt (8325-4); S. aureus Δ hemB ( S. aureus ermB Ω hemB 8325-4, SCV); S. aureus Δ hemB , hemB -complemented ( S. aureus pCX19Ω hemB 8325-4, complemented SCV mutant). Multiplicity of infection (MOI): 20. * significant difference to control cells (P<0.05).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Control, Infection, Incubation, Marker, Western Blot, Negative Control, Mutagenesis

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: (A) HIF 1α immunohistochemistry and (B) hematoxilin-eosin staining of renal mouse abscesses induced by intravenous infection with S. aureus seven days upon infection (following sections). HIF 1α positive nuclei are most abundant within the abscesses borders, some interstitial staining also occurs within the renal parenchyma. In addition, renal tubules in the vicinity of the abscess stain also positively (halfed arrow in B). A: arcuate artery, G: glomerulus, V: arcuate vein; arrowheads: margin of abscesses. Magnifications: (A) 250×, (B) 100×.

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Immunohistochemistry, Staining, Infection

Journal: PLoS ONE

Article Title: Activation of Hypoxia Inducible Factor 1 Is a General Phenomenon in Infections with Human Pathogens

doi: 10.1371/journal.pone.0011576

Figure Lengend Snippet: (A) Survival of NMRI mice after intraperitoneal infection with S. aureus (wt; S. aureus 8325-4; n = 8) or S. aureus Δ hemB (SCV, S. aureus erm Ω hemB 8325-4; n = 8). Note the higher susceptibility of mice infected with S. aureus 8325-4 compared to the mice infected with S. aureus Δ hemB . *significant difference: P<0.001. (B) Inhibition of DFO induced HIF-1 activation in HeLa-229 cells by 17-DMAG. Cells were incubated with 17 DMAG (10 µmol/L) for 16 hours following induction of HIF 1 activation by the iron chelator DFO (200 µmol/L) for six hours. HIF 1α protein was analyzed in Western blots (loading control: actin). Negative control: uninfected cells. (C) Survival rate of NMRI mice after intraperitoneal infection with S. aureus ( S. aureus 8325-4). One group of mice was treated 24 h and 16 h before infection with the HIF-1 inhibiting compound 17-DMAG (25 mg/g body weight). Note the higher survival rate of 17-DMAG-treated mice (n = 45) compared with control mice (n = 45). *significant difference: P = 0.005 (Kaplan-Meier analysis).

Article Snippet: Peritoneal HIF-1α activation was analyzed by immunoblotting of the shock-frozen peritoneal specimens using the Nuclear Extract Kit (Active Motif; Rixensart, Belgium), a mouse monoclonal anti-HIF-1α (H1α67) antibody (Novus Biologicals, Littleton, USA) and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibodies (Dako).

Techniques: Infection, Inhibition, Activation Assay, Incubation, Western Blot, Control, Negative Control