Journal: Aging and Disease

Article Title: Human Umbilical Cord Blood Cells Have Trophic Effects on Young and Aging Hippocampal Neurons in Vitro

doi:

Figure Lengend Snippet: Expression of neuron related antigens and human antigen in the co-culture of aging hippocampal neurons and HUCB cells after 14 DIV.A) Numerous human mitochondria positive cells (green, arrows) were scattered and around MAP2+ aging hippocampal neurons. B) The human mitochondria positive cells (green, arrowhead) did not co-express the proliferative marker BrdU (red, arrows). C) Most of the aging hippocampal cells were positive for VEGFR1 antigen (red, arrow) but there was no co-expression of VEGFR1 with the immature marker, Nestin (green). D) Similar to VEGFR1, a large number of aging hippocampal cells expressed antigen for VEGFR2 (red, arrows) and some of these VEGFR2+ cells also expressed GABAAr (green, arrowheads). E) Numerous cells were detected that expressed antigen to EAAC1 (green, arrows) and a few cells were observed expressing TH (F; green, arrows) and Synaptophysin (G and H; green, arrows). I) Many GFAP+ cells (green) were found in aging hippocampal cultures. Scale bar = 50 μm in (A, E, F), 20 μm in (B–D, G, H), and 100 μm in (I).

Article Snippet: They were then incubated with the primary antibody: polyclonal rabbit anti-microtubule-associated protein 2 (MAP2) antibody (1:1000, Chemicon), polyclonal rabbit anti-glial fibrillary acidic protein (GFAP) antibody (1:500, Dako), chicken anti-GFAP polyclonal antibody (1:400, Chemicon [Millipore]), mouse anti GFAP antibody (1: 1000), mouse anti-human mitochondria (1:50, Chemicon, [Millipore]), neuronal class III β-Tubulin (TuJ1) polyclonal antibody (1:2000, Covance), mouse anti-Nestin antibody (1:100, BD Biosciences Pharmingen), mouse anti-synaptophysin monoclonal antibody (2μg/ml; Chemicon [Millipore]), mouse anti-Tyrosine hydroxylase (TH, 1:100; Chemicon [Millipore]), mouse anti-GABA A receptor (GABAAr), β-chain monoclonal antibody (20 μg/ml; Chemicon [Millipore]), goat anti-glutamate transporter (EAAC1) polyclonal antibody (1:4,000, Chemicon [Millipore]), rabbit polyclonal antibody to VEGF receptor 1(VEGFR1, 1: 100; Abcam), rabbit polyclonal antibody to VEGF receptor 2 (VEGFR2, 1:50; Abcam) at 4°C for 24 hours.

Techniques: Expressing, Co-Culture Assay, Marker

Journal: Aging and Disease

Article Title: Human Umbilical Cord Blood Cells Have Trophic Effects on Young and Aging Hippocampal Neurons in Vitro

doi:

Figure Lengend Snippet: Relative Expression of Neural Antigens by Cultured Hippocampal Neurons

Article Snippet: They were then incubated with the primary antibody: polyclonal rabbit anti-microtubule-associated protein 2 (MAP2) antibody (1:1000, Chemicon), polyclonal rabbit anti-glial fibrillary acidic protein (GFAP) antibody (1:500, Dako), chicken anti-GFAP polyclonal antibody (1:400, Chemicon [Millipore]), mouse anti GFAP antibody (1: 1000), mouse anti-human mitochondria (1:50, Chemicon, [Millipore]), neuronal class III β-Tubulin (TuJ1) polyclonal antibody (1:2000, Covance), mouse anti-Nestin antibody (1:100, BD Biosciences Pharmingen), mouse anti-synaptophysin monoclonal antibody (2μg/ml; Chemicon [Millipore]), mouse anti-Tyrosine hydroxylase (TH, 1:100; Chemicon [Millipore]), mouse anti-GABA A receptor (GABAAr), β-chain monoclonal antibody (20 μg/ml; Chemicon [Millipore]), goat anti-glutamate transporter (EAAC1) polyclonal antibody (1:4,000, Chemicon [Millipore]), rabbit polyclonal antibody to VEGF receptor 1(VEGFR1, 1: 100; Abcam), rabbit polyclonal antibody to VEGF receptor 2 (VEGFR2, 1:50; Abcam) at 4°C for 24 hours.

Techniques: Expressing, Cell Culture

Journal: Journal of Neuroinflammation

Article Title: Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation

doi: 10.1186/s12974-020-01833-1

Figure Lengend Snippet: Increased levels of histamine receptor 2 (HR2) measured in the intestinal samples of young and aged mice for 6-h, 24-h, and 7-days post-stroke compared to the age matched controls. a mRNA expression levels of HR2 quantified by qPCR in young and aged mice at 6 and 24 h post-stroke. b (i) Protein levels of HR2 quantified by IHC followed by image J quantification in aged mice at 7 days post-stroke and c mRNA levels of HR2 quantified by RNA in situ hybridization and analyzed by image-J in the intestine of Ag mice at 7 days post-stroke. n = 3 (sham) and 5 (stroke) per group ( a ) and 5 (sham) and 5 (stroke) per group ( b and c ). Data are expressed as mean ± SEM ( a ), ( b ), and ( c ), as well as individual values. * P < 0.05, ** P < 0.01, *** P < 0.001. P values were calculated using one-way analysis of variance with Tukey multiple comparisons correction ( a ) or with two-tailed unpaired Student’s t test ( b and c ). b Brown-HR2 protein; blue-Nuclei ( b ) purple dots (red arrows)-mRNA HR2

Article Snippet: Formalin fixed, paraffin-embedded intestinal (cecal, ileum) tissue sections (4 μm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens (1) histamine receptor 2 (AHR-002, Alomone labs, Israel), (2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer’s instructions.

Techniques: Expressing, RNA In Situ Hybridization, Two Tailed Test