Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Degradation of transferrin by A. fumigatus in liquid cultures. A. fumigatus was incubated in MEM containing 10% human serum (A) or 2.5 μM holotransferrin (B). Supernatants were withdrawn from the cultures after the number of hours indicated above the lanes, and the presence of transferrin was determined by Western blotting following sodium dodecyl sulfate-PAGE. Controls (lanes C) were uninoculated samples incubated for 286 h. The band underneath transferrin in panel A is another protein that cross-reacted with the polyclonal antitransferrin antibody.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation, Western Blot

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Iron removal from transferrin by A. fumigatus. A. fumigatus was cultured in MEM containing 2.5 μM purified human holotransferrin (A) or 10% human serum (B). Culture media were withdrawn, and the iron saturation of transferrin was analyzed by urea-PAGE. Transferrin was visualized by Western blotting. The numbers above the lanes represent the hours of incubation with A. fumigatus. Fe2-Tf, holotransferrin; Apo-Tf, apotransferrin.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Cell Culture, Purification, Western Blot, Incubation

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: A. fumigatus can transport iron from transferrin across a dialysis membrane. A. fumigatus was inoculated into MEM in which a dialysis bag containing holotransferrin (25 μM) was suspended. A. fumigatus was incubated for 48 h, and then transferrin was withdrawn from the dialysis bag and analyzed by urea-PAGE (lane +). An uninoculated control flask containing MEM plus transferrin in a dialysis bag also was examined (lane −). Pure holotransferrin (Fe2-Tf) and apotransferrin (Apo-Tf) standards also were run.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Iron saturation of transferrin following incubation with A. fumigatus siderophores. Purified desferrisiderophores were incubated with holotransferrin (25 μM) at 37°C for 16 h. Desferriferrichrome, desferriferricrocin, and desferritriacetylfusarinine C were serially diluted to final concentrations of 5 μM (lanes 1), 50 μM (lanes 2), 500 μM (lanes 3), and 5 mM (lanes 4). Controls containing holotransferrin alone also were run (lanes 0). holo-Tf, holotransferrin; Fe-Tf, monoferric transferrin; apo-Tf, apotransferrin.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation, Purification

Journal: Blood

Article Title: Effects of intravenous immunoglobulin on platelet count and antiplatelet antibody disposition in a rat model of immune thrombocytopenia

doi: 10.1182/blood.v100.6.2087

Figure Lengend Snippet: Figure 3. IVIG does not directly bind 7E3. 7E3 (f) (or control IgG, ) and IVIG were combined in vitro, at a constant IVIG concentration (25 mg/mL) and varying 7E3 concentrations (0-0.1 mg/mL). The positive control was a mouse antihuman IgG. Samples were then added to a microplate coated with antihuman IgG. Murine IgG binding was visualized using a secondary antimouse IgG–alkaline phosphatase conjugate. p-Nitrophenyl phosphate was added, and the plates were read at 405 nm (kinetic assay, over 10 minutes). Assay response to 7E3 did not differ from control (P .164), whereas the positive control differed significantly from control (P .001).

Article Snippet: Goat antihuman IgG (no cross-reactivity to goat and mouse serum proteins) and alkaline phosphatase–conjugated goat antimouse IgG (no cross-reactivity to goat and human serum proteins) were both obtained from Rockland (Gilbertsville, PA).

Techniques: Control, In Vitro, Concentration Assay, Positive Control, Binding Assay, Kinetic Assay

Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 2 | Mid-old specific gene expression in old-aged tissues. a–d Representative images of p21Waf1, SAA1, IL1β and SLIT2 IHC analysis in young and old-aged human colon and lung tissues are shown. “1” and “2” indicate high-magnification views of the original figure. Image “1” primarily depicts the epithelium and smooth muscle region, while image “2” depicts a stromal-rich region. Black arrows indicate stained cells in the stromal region. Yellow arrow in (d) indicate SLIT2-positive cells in

Article Snippet: Human SAA1 concentration fromplasmobtained fromAjouUniversityHospital, was measured using a Human Serum Amyloid A1 DuoSet ELISA (DY301905, R&D systems).

Techniques: Gene Expression, Staining

Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 3 | The Effect of SAA1 on old-aged tissues. a Schematic drawing of the effect of SAA1 on tissue. b Young fibroblasts or c smooth muscle cells (PASMCs) were treated with rhSAA1 for 24 h and SASP expression was analyzed using real-time PCR (left panel). MMP9 protein expression was analyzed by ELISA (right panel). d Serially dissected human colon tissues were subjected to IHC analysis for SAA1 and MMP9. Corresponding quantification data is shown in the right panel. IHC results were presented as the percentage of positive cells in the stromal region. e PASMCs were treated with rhSAA1 at the indicated concentration for 24 h and real-time PCR was performed using primers for atrophy-related genes. f IHC ana- lysis for SAA1 in colon tissue from young and elderly subjects, and muscular mucosa thickness was measured. g IHC analysis for type IV collagen of colon tissues from young and elderly subjects is shown. “1” and “2” indicate high-magnification views of the original figure (left panel). Each protein expression was presented as weak, moderate, and strong (right upper panel). mRNA expression level of COL4A1 and COL4A2 in fibroblasts/smooth muscle cells was analyzed between young and old subjects in scRNA-seq data set (GSE178341)34 (right lower panel). h COL4A1,

Article Snippet: Human SAA1 concentration fromplasmobtained fromAjouUniversityHospital, was measured using a Human Serum Amyloid A1 DuoSet ELISA (DY301905, R&D systems).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Concentration Assay, Lysis

Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 6 | SLIT2 has an impact on mid-old cells. a Mid-old cells were treated for 24 h with rhSLIT2, and inflammatory gene expression was analyzed using real-time PCR (upper panel). IL1β and SAA1 protein level was analyzed by ELISA in rhSLIT2-treated mid-old cells (lower panel). b Mid-old cells were treated with rhSLIT2 for the indi- cated times and analyzed for Pyk2-NFκB signaling by western blot analysis (left panel). The number of nuclear p-NFκB-positive cells was quantified in mid-old cells using IF staining (right panel). c SLIT2-expressing lentivirus infected into mid-old fibroblasts, and inflammation related genes expression including SAA1 and IL1β were analyzed using real-time PCR (right lower panel). IL1β and SAA1 protein level was analyzed by ELISA in SLIT2-overexpressing mid-old cells (left lower panel). d The morphology and cell size of mid-old cells were analyzed after treating rhSLIT2 for 20 days. Cell growth rate was analyzed every 4 days for 20 days by counting the number of cells. e The expression of p53 and p21Waf1 were measured in mid-old cells after administration of rhSLIT2 at the indicated concentration for 2 days using real-time PCR (upper panel) and western blot analysis (lower panel). f The changes in gene expression of p53 and p21Waf1-regulating genes were eval- uated after treating cells with rhSLIT2 using RNA-seq FPKM count (left panel). The mRNA expression of SOX2 and OCT4 were analyzed using real-time PCR (right

Article Snippet: Human SAA1 concentration fromplasmobtained fromAjouUniversityHospital, was measured using a Human Serum Amyloid A1 DuoSet ELISA (DY301905, R&D systems).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Staining, Expressing, Infection, Concentration Assay, RNA Sequencing