Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: L1 protein associated with condensed chromosomes retains the reactivity of the H16.V5 epitope. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without Click-iT reaction buffer without dye. Next, the cells were incubated with L1 conformationally dependent mouse MAb H16.V5 antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: L1 protein associated with condensed chromosomes retains the reactivity of the H16.56E epitope. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without Click-iT reaction buffer without dye. Next, the cells were incubated with L1 conformationally dependent mouse MAb H16.56E antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: The linear epitope of the 33L1-7 antibody is hidden in the L1 protein associated with condensed chromosomes. (A and B) At 24 hpi, HaCaT cells infected with EdU-labeled pseudovirus were fixed, permeabilized, and treated with or without with Click-iT reaction buffer without dye. Next, the cells were incubated with mouse MAb 33L1-7 antibody (green) and AF488-conjugated anti-α-tubulin (white). Lastly, the cells were treated with Click-iT reaction buffer with AF555 dye (red) to stain the EdU-labeled pseudogenome and mounted in DAPI (blue). Note that colocalization of the EdU puncta and L1 protein is denoted as a yellow color.

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Papillomavirus Major Capsid Protein L1 Remains Associated with the Incoming Viral Genome throughout the Entry Process

doi: 10.1128/JVI.00537-17

Figure Lengend Snippet: Some full-length L1 protein accompanies the viral genome to the nucleus. (A) HeLa cells were infected with HPV16 pseudovirus with or without 1.5 μM Eg5 inhibitor III (Eg5i) and tracked via live-cell imaging using the IncuCyte Zoom for 48 h. Note that the images are depicted at 18 hpi. (B) HaCaT cells were infected with EdU-labeled HPV16 pseudovirus in the presence of 1.5 μM Eg5i. The cells were fixed at 24 hpi and permeabilized with either digitonin at 0.625 μg/ml or 0.5% TX-100 and then treated with AF555 (green) in Click-iT reaction buffer. The cells were permeabilized again with 0.5% TX-100 and treated with AF647 (red) in Click-iT reaction buffer. Lastly, the cells were mounted in DAPI (blue). (C) HaCaT cells were infected with EdU-labeled pseudovirus and treated with 1.5 μM Eg5i. At 24 hpi, the cells were fixed and permeabilized in 0.5% TX-100. Next, the cells were treated with AF555 (red) in Click-iT reaction buffer, followed by incubation with AF488-conjugated anti-α-tubulin (white) and MAb 33L1-7 (green). Lastly, the cells were mounted in DAPI (blue). Note the colocalization between EdU and L1 signal denoted by white arrows. (D) HeLa cells were infected with HPV16 pseudovirus in the presence of 1.5 μM Eg5i. Cells were trypsinized for 2 min, and monoastral cells were collected. Next, the cells were treated with 15 μl of 0.25% trypsin for 1 h at 37°C. The cells were lysed by passage through a 1-ml syringe with a 25-gauge needle 40 times. Cell lysates were incubated for 1 h at 37°C once more, the trypsin was inactivated, and the samples were analyzed by Western blot analysis with a cocktail of HPV16 L1-specific mouse MAbs (IID5, 33L1-7, and 312F).

Article Snippet: The microtubule network was detected using mouse MAb anti-α-tubulin conjugated to Alexa Fluor 488 (AF488; catalog no. 8058S; Cell Signaling).

Techniques: Infection, Live Cell Imaging, Labeling, Incubation, Western Blot

Journal: European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery

Article Title: Decellularized mitral valve in a long-term sheep model.

doi: 10.1093/ejcts/ezx485

Figure Lengend Snippet: Figure 7: Immunostaining of the decellularized mitral valve (DMV) and the biological mitral valve (BMV). (A and B) DMV and (C and D) BMV. (A, C) CD45 and DNA; (B, D) procollagen I and DNA. DMV shows collagen-producing cells within the leaflets. BMV has almost no cells, positive for procollagen I. The CD45-positive cells in the BMV tend to form nodules (picture of the region, close to the annulus). BMV has only several CD45-positive cells on the surface. Ao: Aorta ascendens.

Article Snippet: D ow nloaded from https://academ ic.oup.com /ejcts/article/53/6/1165/4828177 by guest on 22 M ay 2024 • Collagen IV (clone CI22, DAKO, Hamburg, Germany) • Collagen I C2345 clone COL-I (Sigma-Aldrich Chemie GmbH, Munich, Germany) • CD31 [monoclonal mouse immunoglobulin G (IgG)2a, Serotec, Raleigh, NC, USA] • von Willebrand factor (polyclonal rabbit IgG, DAKO) • Endothelial nitric oxide synthase (monoclonal mouse IgG1, eNOS/NOS Type III, BD Transduction Laboratories, San Jose, CA, USA) • CD45 [mouse monoclonal (clone OX-1) IgG, BioRad, Oxford, UK] • Procollagen (monoclonal mouse IgG1, M-38-c, Developmental Studies Hybridoma Bank, Iowa City, IA, USA) The images were obtained using the microscope AxioObserver A1, with the camera AxioCamMRm (Carl Zeiss, Jena, Germany).

Techniques: Immunostaining

Journal: Medicine

Article Title: Circulating Tumor Cell Analyses in Patients With Esophageal Squamous Cell Carcinoma Using Epithelial Marker-Dependent and -Independent Approaches

doi: 10.1097/MD.0000000000001565

Figure Lengend Snippet: Within CellSearch TM system, CTCs are defined as epithelial origin cells with round or oval morphology, cell size >4 μm in diameter, cell surface antigen EpCAM, CK, and DAPI positive, CD45 antigen negative (EpCAM+/CK+/DAPI+/CD45−).

Article Snippet: The slides were then incubated overnight at 4°C in a wet chamber, with anti-CD45 Rat mouse monoclonal antibody (Santa-Cruz Biotechnology, Inc., Dallas, Texas, USA), anti-CK8/18/19 antibody (Abcam Trading (Shanghai) Company Ltd., Pudong, Shanghai, China), and antivimentin antibody (Abcam).

Techniques:

Journal: Medicine

Article Title: Circulating Tumor Cell Analyses in Patients With Esophageal Squamous Cell Carcinoma Using Epithelial Marker-Dependent and -Independent Approaches

doi: 10.1097/MD.0000000000001565

Figure Lengend Snippet: Immunofluorescent staining for characterization of CTCs/CTM detected by ISET assay. (A–D) Immunofluorescent staining characterization of CTC/CTM detected by ISET assay. Stained with Hoechst (blue fluorescence for nucleus), CK8/18/19 (green fluorescence for epithelial cells), vimentin (purple fluorescence for EMT cells), and CD45 (red fluorescence for leukocytes). (A) Cell line control; (B) CTC: expression of CK-positive in their CTCs (CK+/Vimentin−/CD45−); (C) CTM: expression of CK-weakly positive and Vimentin-positive in cells within CTM (CK+/Vimentin+/CD45−); (D) EMT-CTM: heterogeneity was observed for CK-negative and Vimentin-positive (CK−/Vimentin+/CD45−). The cells were analyzed under ×40 magnification.

Article Snippet: The slides were then incubated overnight at 4°C in a wet chamber, with anti-CD45 Rat mouse monoclonal antibody (Santa-Cruz Biotechnology, Inc., Dallas, Texas, USA), anti-CK8/18/19 antibody (Abcam Trading (Shanghai) Company Ltd., Pudong, Shanghai, China), and antivimentin antibody (Abcam).

Techniques: Staining, Fluorescence, Control, Expressing