Journal: Journal of neuroscience methods

Article Title: A white matter stroke model in the mouse: Axonal damage, progenitor responses and MRI correlates

doi: 10.1016/j.jneumeth.2009.03.017

Figure Lengend Snippet: Cell death in white matter one day after stroke. (A) Immunohistochemical staining for AIF in contralateral hemisphere. (B) AIF stain in the hemisphere ipsilateral to stroke. Note positive cells in white matter and deepest cortical layers, and linear rows of AIF immunoreactive cells (arrows). (C) CAII stain for oligodendrocytes (green) activated PARP immunoreactivity (red). Note multiple CAII and activated PARP double positive cells in subcortical white matter (D) In situ nick translation showing double stranded DNA nicking in apoptotic cells in subcortical white matter. (E) Transferrin immunoreactive oligodendrocytes in control subcortical white matter region. (F) Transferrin immunoreactive oligodendrocytes 7 days after subcortical white matter stroke. (G) NG2 immunoreactive OPCs in control subcortical white matter region. (H) NG2 immunoreactive OPCs 14 days after subcortical white matter stroke. ctx = cortex, str = striatum, wm = subcortical white matter. Bar in (B) = 50 μm and applies to (A and B); bar in (Cand D) = 40 μm, Bar in (F) = 20 μm and applies to (E–H) (for interpretation of the references to color in this figure legend, the reader is referred to the web version of the article).

Article Snippet: Primary antibodies were: rat anti-myelin basic protein (1:800, Carlsbad, CA), rabbit anti-myelin basic protein (MBP)(1:200, Abcam, Cambridge, MA), mouse anti-NeuN (1:500, Abcam), rat anti-GFAP (1:1000, Invitrogen), sheep anti-carbonic anhydrase II (1:600, Morphosys, Germany), sheep and rat anti-BrdU (1:500, Abcam), rabbit anti-transferrin (1:75, Abcam), mouse anti-apoptosis inducing factor (1:200, AIF, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-activated Poly(ADP-ribose)polymerase 1 (PARP, 1:100, Cell Signaling, Danvers, MA), rabbit anti-NF200 (1:2000, Sigma, St. Louis, MO), mouse anti-SMI31 and anti-SMI-32 (1:1000, Abcam), rabbit anti-NG2 (1:200, Invitrogen); goat anti-transferrin (1:100, MP Biomedicals, Solon, OH), rabbit anti-IBA1 (1:1000, Wako Chemicals, Richmond, VA) and NeuN (1:500).

Techniques: Immunohistochemical staining, Staining, In Situ, Nick Translation

Journal: Journal of neuroscience methods

Article Title: A white matter stroke model in the mouse: Axonal damage, progenitor responses and MRI correlates

doi: 10.1016/j.jneumeth.2009.03.017

Figure Lengend Snippet: Early and late response of OPCs and oligodendrocytes following subcortical stroke. BrdU pulse-chase experiment at day 7 and 28 after subcortical stroke. (A) Representative images of BrdU double labeling of oligodendrocyte progenitor cells (OPC) with OPC marker NG2 (top panel), and oligodendrocytes with specific marker transferrin (bottom panel) at day 7. The insets correspond to higher power views. Bar in right middle panel of A = 20 μm. (B) Total number of double labeled cells estimated by stereology: means ± standard deviations. NG2 7 day vs. 28 day 3918.33 ± 238.03, 2150.0 ± 413.54, p = 0.003; transferrin 7 day vs. 28 day 3665.0 ± 456.54, 1941.67 ± 677.98, p = 0.02.

Article Snippet: Primary antibodies were: rat anti-myelin basic protein (1:800, Carlsbad, CA), rabbit anti-myelin basic protein (MBP)(1:200, Abcam, Cambridge, MA), mouse anti-NeuN (1:500, Abcam), rat anti-GFAP (1:1000, Invitrogen), sheep anti-carbonic anhydrase II (1:600, Morphosys, Germany), sheep and rat anti-BrdU (1:500, Abcam), rabbit anti-transferrin (1:75, Abcam), mouse anti-apoptosis inducing factor (1:200, AIF, Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-activated Poly(ADP-ribose)polymerase 1 (PARP, 1:100, Cell Signaling, Danvers, MA), rabbit anti-NF200 (1:2000, Sigma, St. Louis, MO), mouse anti-SMI31 and anti-SMI-32 (1:1000, Abcam), rabbit anti-NG2 (1:200, Invitrogen); goat anti-transferrin (1:100, MP Biomedicals, Solon, OH), rabbit anti-IBA1 (1:1000, Wako Chemicals, Richmond, VA) and NeuN (1:500).

Techniques: Pulse Chase, Labeling, Marker

Journal: Viruses

Article Title: Nisoldipine Inhibits Influenza A Virus Infection by Interfering with Virus Internalization Process

doi: 10.3390/v14122738

Figure Lengend Snippet: Nisoldipine inhibits endocytosis of IAV. ( A ) Nisoldipine could reduce the absorption of transferrin. A549 cells were pretreated with nisoldipine for 3 h and then incubated with transferrin (15 µg/mL) at 37 °C for 30 min. The cells were lysed with RIPA and then subjected to Western blotting with a rabbit anti-transferrin mAb. ( B ) Nisoldipine could reduce the number of clathrin-encapsulated endocytic vesicles. Scale bar: 500 nm. Magnification: 40,000×. Red arrows point to the clathrin-coated vesicle. ( C ) Nisoldipine could reduce the expression of caveolin-1. ( D ) Transferrin uptake was decreased when the cell was pretreated with nisoldipine. CPZ was used as a positive control. Scale bar: 20 μm. Tf-568 is shown in red. ( E ) The marked reduction of CTB endocytosis was observed in nisoldipine-treated cells. MβCD was used as a positive control. CTB-FITC is visualized in green. Data are presented as mean ± SD (** p < 0.01, *** p < 0.001).

Article Snippet: The following antibodies were used: influenza A virus PB2 protein antibody (Genetex, GTX125926); influenza A virus NP antibody (Genetex, GTX125989); mouse anti-transferrin antibody (Bioss, bsm-33244M); rabbit anti-caveolin-1 antibody (Bioss, bs-1453R); anti-Cav1.2 antibody (Abcam, ab84814); and GAPDH (glyceraldehyde-3-phosphate dehydrogenase)/β-actin rabbit mAb (Bioss, bs-0061R-2).

Techniques: Incubation, Western Blot, Expressing, Positive Control

Journal:

Article Title: Phagocytosis and Intracellular Fate of Aspergillus fumigatus Conidia in Alveolar Macrophages

doi: 10.1128/IAI.71.2.891-903.2003

Figure Lengend Snippet: Maturation of A. fumigatus-containing phagosomes in HAMs. HAMs were incubated with conidia (2:1 ratio of conidia to macrophages) and were fixed at 5, 10, 30, and 60 min after the beginning of phagocytosis. After permeabilization with saponin, cells were labeled at 10 min (a to c and e to g) or 60 min (d and h to p) postingestion with the specific monoclonal or polyclonal anti-TfR (1/1,000), anti-EEA1 (1/500), anti-Rab 7 (1/50), and anti-Lamp1 (1/100) antibodies and secondary antibodies conjugated to Texas Red. (a to c) Arrowheads indicate TfR-positive (a), EEA1-positive (b), and Rab 7-positive (c) staining surrounding the conidia. (d) Cells were labeled at 60 min postingestion, and the arrowhead indicates positive staining for Lamp1. (e to h) Corresponding DIC images. The arrowheads indicate conidia. Note that the conidia outside the cells (thin arrows in panels c, g, j, and n) were not labeled by the same antibodies. (i to k) Lack of labeling with the anti-TfR, anti-EEA1, and anti-Rab7 antibodies. (l) Cells were labeled with control IgG1 MAb. (m to p) Corresponding DIC images. Bar, 2 μm.

Article Snippet: The following primary antibodies were used in this study: a rabbit polyclonal antibody (pAb) specific for A. fumigatus conidia ( 67 ), two anti-transferrin receptor (anti-TfR) monoclonal antibodies (MAbs) (RI7-217 and OKT9; American Type Culture Collection) recognizing the murine or human TfR (kindly provided by A. Dautry, Institut Pasteur), a mouse anti-early endosomal antigen 1 (anti-EEA1; Transduction Laboratories) MAb recognizing the human protein, an anti-Rab7 pAb ( 52 ) (kindly provided by S. Méresse, CIML, Marseille, France), two anti-LAMP1 MAbs recognizing the human and the murine proteins (CD107a; PharMingen), and an anti-human cathepsin D pAb ( 43 ) (kindly provided by W. Gregory, Washington University School of Medicine).

Techniques: Incubation, Labeling, Staining