Journal: The Journal of Neuroscience

Article Title: Protogenin Defines a Transition Stage during Embryonic Neurogenesis and Prevents Precocious Neuronal Differentiation

doi: 10.1523/JNEUROSCI.0473-10.2010

Figure Lengend Snippet: Expression of PRTG protein in the mouse embryo. A, Expression of rat prtg in the nervous system at different stages of development as analyzed by the Northern blot. Ad, Adult; Br, brain; Nt, neural tube. B, Specificity of PRTG antibodies. Cell lysates of HEK293T cells transfected with the control vector, prPRTGf (rat full-length PRTG), prPRTGΔc (rat PRTG without cytoplasmic domain), or prPRTGc (rat cytoplasmic tail), and rat E10.5 neural tube lysate (Embryo) were subjected to Western blot using α-PRTG1 and α-PRTG2 mAbs. Addition of peptide E (E; ectodomain) or peptide C (C; cytoplasmic tail) competes out the bands recognized by the monoclonal antibodies. C, Schematic structure of PRTG. Peptide E or C for generating antibodies is marked in blue. D–H, Transverse sections through the mid-body level of mouse at E7 (D), E7.75 (E), E8.25 (F), E9.5 (G), and E10.5 (H) were stained using α-PRTG2 mAb (red). The dorsal is up in all panels. D, At E7, PRTG labeling is weakly detected in the mesoderm (me), whereas Oct4 is restricted to the ectoderm (ec; green). E, At E7.75, PRTG is detected in neuroepithelium, mesoderm, endoderm, and posterior epiblasts (arrows). The yolk sac (ys) surrounding the embryo is nonspecifically stained by the secondary antibodies. F, G, PRTG protein is present in almost all cells between E8.25 and E9.5 (red) but not in the notochord (n), differentiating cardiac cells, and some mesenchymal cells. The endocardium is labeled with anti-endoglin antibody (green). ac, Atrial chamber; dm, dermomyotome; vc, ventricular chamber; sc, spinal cord. H, PRTG is not detectable at E10.5. I, Expression of PRTG (red) and SSEA1 (green) in a transverse section of E7 and E8.25 mouse embryos. J–N, Expression of PRTG (red) and nestin (J), Ascl1 (K), TuJ1 (L), MAP2 (M), and NeuN (N) (green) in adjacent transverse sections through the thoracic level of E9.5 (top) and E10.5 (bottom) mouse embryos.

Article Snippet: Goat anti-mouse endoglin (1:100) was purchased from R & D Systems.

Techniques: Expressing, Northern Blot, Transfection, Control, Plasmid Preparation, Western Blot, Bioprocessing, Staining, Labeling

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Migrating lymph dendritic cells contain intracellular CD40 that is mobilized to the immunological synapse during interactions with antigen-specific T lymphocytes.

doi: 10.4049/jimmunol.1202010

Figure Lengend Snippet: FIGURE 2. Lymphocyte and DC interaction in SS pseudo-afferent rat lymph does not induce polarization of DC CD40. (A) Pseudo-afferent lymph stained with 7-AAD; (B) pseudo-afferent lymph stained with anti-CD103 FITC; (C) overlay of images (A, B); (D) high intensity image of (C); (E) line contour image of (C). Arrows show points of contact between lymphocytes and DC. (F) CD40 distribution throughout the cell membrane of a pseudo- afferent DC in SS conditions. (G) CD40 distribution remains throughout the cell membrane during SS conditions even when DC interacts with lymphocyte. (H) The 90˚ reconstructed image of pseudo-afferent DC closely interacting with at least two lymphocytes during SS conditions. CD40 still remains throughout the cell membrane and is not polarized toward the lymphocytes. Scale bars at bottom left of images (A) and (F), 10 mm.

Article Snippet: The cells were then incubated with goat anti-mouse CD40 (T-20 Ab; 1:100; Santa Cruz Biotechnology), followed by rabbit anti-goat IgG conjugated to FITC (1:400; Serotec) and mouse anti-rat CD3 (1:100; Serotec), followed by goat anti-rat IgM conjugated to tetramethylrhodamine isothiocyanate (1:500; Serotec).

Techniques: Staining, Membrane

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Migrating lymph dendritic cells contain intracellular CD40 that is mobilized to the immunological synapse during interactions with antigen-specific T lymphocytes.

doi: 10.4049/jimmunol.1202010

Figure Lengend Snippet: FIGURE 3. Freshly isolated SS and LPS-induced LDC show intracellular expression of CD40. FACS histograms showing density-enriched SS and LPS- induced LDC were double immuno- labeled using anti-CD103 biotin/strep- tavidin-PE and L-17 (anti-CD40) poly- clonal Ab. For surface labeling (A), cells remained unfixed and nonpermeabilized throughout, and for intracytoplasmic la- beling (B), cells were fixed and per- meabilized prior to CD40 labeling. CD40 expression by SS LDC and LPS- induced LDC is shown by the filled area, and isotype controls are shown by the dashed line. Cells were either unstimu- lated (steady state) (A, B) or stimulated by LPS (C, D). Each plot is representa- tive of three independent experiments.

Article Snippet: The cells were then incubated with goat anti-mouse CD40 (T-20 Ab; 1:100; Santa Cruz Biotechnology), followed by rabbit anti-goat IgG conjugated to FITC (1:400; Serotec) and mouse anti-rat CD3 (1:100; Serotec), followed by goat anti-rat IgM conjugated to tetramethylrhodamine isothiocyanate (1:500; Serotec).

Techniques: Isolation, Expressing, Labeling

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Migrating lymph dendritic cells contain intracellular CD40 that is mobilized to the immunological synapse during interactions with antigen-specific T lymphocytes.

doi: 10.4049/jimmunol.1202010

Figure Lengend Snippet: FIGURE 4. Freshly isolated SS and LPS-induced LDC express multiple protein isoforms of CD40. Western blots show CD40 protein isolated from den- sity-enriched SS and LPS-induced LDC. Whole-cell lysates (protein equivalent of 5 3 106 LDC) were electrophoretically separated prior to transfer onto a nitrocellulose membrane. CD40 immunoreactivity was detected using anti-goat HRP-conjugated second- ary Ab prior to peroxidase development using ECL with an exposure time of 5 min. (A) Blots probed with L-17 CD40 polyclonal Ab; (B) blots probed with type I CD40 isoform-specific Ab (T-20); (C) blots probed with T-20 Ab that had been preadsorbed with CD40 block- ing peptide.

Article Snippet: The cells were then incubated with goat anti-mouse CD40 (T-20 Ab; 1:100; Santa Cruz Biotechnology), followed by rabbit anti-goat IgG conjugated to FITC (1:400; Serotec) and mouse anti-rat CD3 (1:100; Serotec), followed by goat anti-rat IgM conjugated to tetramethylrhodamine isothiocyanate (1:500; Serotec).

Techniques: Isolation, Western Blot, Membrane, Blocking Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Migrating lymph dendritic cells contain intracellular CD40 that is mobilized to the immunological synapse during interactions with antigen-specific T lymphocytes.

doi: 10.4049/jimmunol.1202010

Figure Lengend Snippet: FIGURE 6. Polarization of CD40 to the IS in rat pseudo-afferent LDC occurs only after at least 3-h culture with allogeneic CD4+ lymphocytes and is absent after 24 h. Confocal images of CD11c+ LDC and CD4+ T lymphocyte cultures shown in Fig. 5. (A, E, and I) CD3/tetramethylrhodamine isothiocyanate immunolabeling of LDC/T lymphocyte clusters; (B, F, and J) CD40/FITC immunolabeling of LDC/T lymphocyte clusters and (C, G, and K) overlay of previous images. (D, H, and L) Transmitted light images of the same LDC/T lymphocyte clusters. CD40 polarization to the IS occurs after 6-h culture (E–G), but is absent after 24 h (I–K). Images shown are representative of images obtained from five slides on three separate occasions. Scale bars at bottom left of images (A), (E), and (I), 10 mm.

Article Snippet: The cells were then incubated with goat anti-mouse CD40 (T-20 Ab; 1:100; Santa Cruz Biotechnology), followed by rabbit anti-goat IgG conjugated to FITC (1:400; Serotec) and mouse anti-rat CD3 (1:100; Serotec), followed by goat anti-rat IgM conjugated to tetramethylrhodamine isothiocyanate (1:500; Serotec).

Techniques: Immunolabeling

Journal: Apoptosis

Article Title: Adipose stromal/stem cells assist fat transplantation reducing necrosis and increasing graft performance

doi: 10.1007/s10495-013-0878-7

Figure Lengend Snippet: Antibodies for rabbit ASC characterization by FACS

Article Snippet: CD105 , Mouse , SN6 , 1/20 , Serotec , No.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: MHC class II expression and potential antigen-presenting cells in the retina during experimental autoimmune uveitis

doi: 10.1186/s12974-017-0915-5

Figure Lengend Snippet: MHC class II is expressed at the level of the RPE layer and by cells infiltrating the vitreous. Three weeks after adoptive transfer, EAU eye cryosections were prepared and stained for MHC class II ( green ), IBA1 ( red ), or endoglin ( magenta ) detection. Cell nuclei were stained with Hoechst ( blue ). Each picture was chosen as representative of an experiment conducted on three or more animals. The DIC image was added to localize the RPE more precisely. a MHC class II and endoglin expression in the RPE region. Thin arrows point to the RPE layer, and thick arrows indicate choroidal vessels stained with endoglin. b MHC class II and IBA1 expression in the RPE region and corresponding DIC image. Thin arrows point to the RPE layer, and thick arrows indicate the choroid. c MHC class II and IBA1 expression by cells infiltrating the vitreous. Thin arrows indicate cells positive for both markers, and thick arrows point to cells positive for only one marker

Article Snippet: Whole retinas were fixed in 70% ethanol for 1 h, rinsed three times in PBS (10 min each), blocked with a solution containing 3% milk and 3% bovine serum albumin in PBS for 1 h and incubated with anti-MHC class II (rat, 1/200; BD Biosciences, Erembodegem, Belgium) and anti-endoglin (goat, 1/200; BD Biosciences) antibodies in a PBS-BSA1% solution overnight at room temperature.

Techniques: Adoptive Transfer Assay, Staining, Expressing, Marker

Journal: Biomaterials

Article Title: Quantum dots-based in situ molecular imaging of dynamic changes of collagen IV during cancer invasion.

doi: 10.1016/j.biomaterials.2013.07.069

Figure Lengend Snippet: Fig. 3. Dynamic changes of tumor microenvironment in well-differentiated tubular adenocarcinoma of GC. (A) shows collagen IV and macrophages in peri-tumor tissue of GC. (B) shows GC nests are surrounded by intact and dense collagen IV almost without degradation and ECM deposition. (C) shows collagen IV aligned with GC nests is degraded at invasion front (Red arrows) with increased irregular collagen IV deposition in ECM. (DeF) show increased deposition of collagen IV in tumor stroma, presenting stiff and rigid. (GeH) show CD105 expression in peri-tumor tissue and GC tissue, respectively. (I) shows double-staining of macrophages and CD105 in GC tissue, in which tumor macrophages aggregate. (JeL) show multiplexed imaging of collagen IV, macrophages and CD105, in which cancer cells, macrophages and tumor neo-vessels constitute “invasion unit” (Red square indicates an “invasion unit”). Scale bar: 50 mm for A, D, E, G, H & J; 20 mm for B, C, F & K; 10 mm for I & L.

Article Snippet: After blocked with 2% bovine serum albumin (BSA), the slides were first incubated with primary antibodies for rabbit anti-human polyclonal antibody against collagen IV (ab6586, Abcam, England, dilution 1/300), mouse anti-human monoclonal antibody against macrophages (MA1-38069, ABR, USA, dilution 1/300) and goat anti-human polyclonal antibody against CD105 (sc-20072, Santa Cruz, USA, dilution 1/500) for 2 h at 37 C, then with corresponding secondary antibody, including peroxidase labeled anti-goat IgG (H þ L) antibody (14-13-06, KPL, USA, dilution 1/300), peroxidase labeled anti-mouse IgG (H þ L) antibody (074-1806, KPL, USA, dilution 1/300) and peroxidase labeled anti-rabbit IgG (Hþ L) antibody (074-1506, KPL, USA, dilution 1/300) for 30 min at 37 C, respectively.

Techniques: Expressing, Double Staining, Imaging