Journal: Cells

Article Title: Aged Mouse Hippocampus Exhibits Signs of Chronic Hypoxia and an Impaired HIF-Controlled Response to Acute Hypoxic Exposures

doi: 10.3390/cells11030423

Figure Lengend Snippet: Genes of interest transcribed by HIF-α isoforms. HIF-1α transcribes metabolic switching genes lactose dehydrogenase- A ( LDHA ), phosphoglycerate kinase 1 ( PGK1 ), pyruvate dehydrogenase kinase 1 ( PDK1 ), and switched cyclooxygenase 4 ( COX4 ) from isoform 1 to isoform 2. Both HIF-1α and HIF-2α have been reported to transcribe glucose transporter 1 ( GLUT1 ), monocarboxylate transporters ( MCT ) 1 and 4, erythropoietin ( EPO ), and vascular endothelial growth factor ( VEGF ) in a cell-specific manner. HIF-2α is particularly involved in modulating the EPO signal. Glu = glucose; 1,3-BPG = 1,3-biphosphoglycerate; 3-PG = 3-phosphoglycerate; Py = pyruvate; Lac = lactate; TCA = tricarboxylic acid cycle; I, II, III, and IV = complexes I, II, III, and IV of electron transport chain.

Article Snippet: Each sample was incubated with primary antibodies directly conjugated to their corresponding fluorophore, targeting either HIF-1α-dy650 (1:200, NB100-479C) or HIF-2α-AF647 (1:100, NB100-122AF647; Novus Biologicals, Centennial, CO, USA).

Techniques:

Journal: Cells

Article Title: Aged Mouse Hippocampus Exhibits Signs of Chronic Hypoxia and an Impaired HIF-Controlled Response to Acute Hypoxic Exposures

doi: 10.3390/cells11030423

Figure Lengend Snippet: Hippocampal HIF-α protein levels are elevated with age. ( A , E ) HIF-1α ( A ) and HIF-2α ( E ) protein reported as intensity/μm 2 using IHC on hippocampal slices from 3 mo, 9 mo, 18 mo, and 24 mo age groups. ( B , F ) Representative images of HIF-1α ( B ) and HIF-2α ( F ) intensity IHC observations. White spots are HIF-1α, and blue spots are DAPI. ( C , G ) HIF-1α ( C ) and HIF-2α ( G ) protein measured as intensity/μm 2 within hippocampal subregions (dentate gyrus [DG], CA1, and CA3) of different aged mice. ( D , H ) Representative images of HIF-1α ( D ) and HIF-2α ( H ) intensity within each subregion. Images are displayed as merged channels with HIF-1α or HIF-2α in white and DAPI in blue. Scale bar = 150 μm; p ≤ 0.05; * = compared to 3 mo normoxic control; # = compared to 9 mo; ŧ = compared to 18 mo; ϕ = compared to other age-matched regions; small dots = female mice; large dots = male mice.

Article Snippet: Each sample was incubated with primary antibodies directly conjugated to their corresponding fluorophore, targeting either HIF-1α-dy650 (1:200, NB100-479C) or HIF-2α-AF647 (1:100, NB100-122AF647; Novus Biologicals, Centennial, CO, USA).

Techniques: Control

Journal: Cells

Article Title: Aged Mouse Hippocampus Exhibits Signs of Chronic Hypoxia and an Impaired HIF-Controlled Response to Acute Hypoxic Exposures

doi: 10.3390/cells11030423

Figure Lengend Snippet: Hippocampal HIF-α protein levels associated with hypoxia. Comparison of HIF-1α and HIF-2α levels in the hippocampus of male (large dot) and female (small dot) mice exposed to room air (normoxic; open circles) or 3% oxygen for 3 h (hypoxia; filled circles). ( A , E ) HIF-1α ( A ) and HIF-2α ( E ) mean intensity/μm 2 in total hippocampus of different aged mice exposed to room air or hypoxia. ( B , F ) Representative images of HIF-1α ( B ) or HIF-2α ( F ) (illustrated in white) merged with DAPI (blue) in the hippocampus of different aged mice exposed to room air or hypoxia. ( C , G ) HIF-1α ( C ) or HIF-2α ( G ) protein measured as intensity/μm 2 within hippocampal subregions (CA3, CA1, and dentate gyrus [DG]) of different aged mice exposed to room air or hypoxia. ( D , H ) mRNA levels of HIF-1α ( D ) or HIF-2α ( H ) measured by fISH in sections from the same aged mice exposed to room air or hypoxia. Results are reported as the number of spots counted per μm 2 . Scale bar = 150 μm; p ≤ 0.05; * = compared to 3 mo normoxic control; # = compared to 9 mo; ŧ = compared to 18 mo; § = interaction between age and hypoxia.

Article Snippet: Each sample was incubated with primary antibodies directly conjugated to their corresponding fluorophore, targeting either HIF-1α-dy650 (1:200, NB100-479C) or HIF-2α-AF647 (1:100, NB100-122AF647; Novus Biologicals, Centennial, CO, USA).

Techniques: Comparison, Control