Journal: Emerging Infectious Diseases

Article Title: Livestock Susceptibility to Infection with Middle East Respiratory Syndrome Coronavirus

doi: 10.3201/eid2302.161239

Figure Lengend Snippet: Presence of MERS-CoV receptor DPP4 (IHC) and of mucosubstances (PAS) in upper and lower respiratory tract tissues from sheep, pigs, llamas, and horses. A) In the nose, DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of pigs, llamas, and horses but not sheep. PAS staining (magenta) demonstrated more mucous cells in the lining epithelium of sheep and horses and a layer of mucus on the lining epithelium of the horses. B) DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of the trachea, bronchus/bronchioles, and alveoli in the pigs, llamas and horses but not in the sheep. Original magnification ×400 for all images. DPP4, dipeptidyl peptidase-4; IHC, immunohistochemistry; MERS-CoV, Middle East respiratory syndrome coronavirus; PAS, periodic acid–Schiff; term., terminal.

Article Snippet: In brief, we performed DPP4 IHC staining by using 5 μg/mL of polyclonal goat IgG anti-human DPP4 antibody (R&D Systems, Abingdon, UK) and peroxidase-labeled rabbit anti-goat IgG (1:200; DAKO; Agilent Technologies Company, Santa Clara, CA, USA) as a secondary antibody.

Techniques: Membrane, Staining, Immunohistochemistry

Journal: Journal of Pediatric Hematology/Oncology

Article Title: Chemotherapy-induced Alteration of SDF-1/CXCR4 Expression in Bone Marrow–derived Mesenchymal Stem Cells From Adolescents and Young Adults With Acute Lymphoblastic Leukemia

doi: 10.1097/mph.0000000000000220

Figure Lengend Snippet: FIGURE 3. Secretion and expression of SDF-1 in BM-MSCs. A, SDF-1 level was measured by enzyme-linked immunosorbent assay. (a) The bone marrow plasma level of SDF-1 in patients with ALL was significantly increased compared with that in healthy donors and ALL-c (*P < 0.05; n = 10). (b), The same difference was found when SDF-1 levels were measured in the supernatants of MSC cultures after incubation for 72 hours. B, Western blotting was performed to quantify total SDF-1 in the lysates of MSCs. SDF-1 expression in the MSCs of ALL was significantly higher than that in the MSCs of healthy donors and ALL-c. C, The difference was also found at the transcriptional level by real-time PCR (*P < 0.05, n = 10). D, Surface-CD26 expression on the MSCs from ALL patients measured by flow cytometry was significantly lower than that in healthy donors and ALL-c (*P < 0.05; n = 10). E, Total CD26 was measured by Western blot in the MSCs. The fold difference was shown among the 3 types of MSCs. F, The same difference was also found at the transcriptional level (*P < 0.05; n = 10). ALL indicates acute lymphoblastic leukemia; BM-MSC, bone marrow–derived mesenchymal stem cell; GADPH, glyceraldehyde- 3-phosphate dehydrogenase; PCR, polymerase chain reaction.

Article Snippet: Membranes were hybridized with the following primary antibodies: rabbit anti-human CXCR4 (1:1000; Abcam), goat antihuman SDF-1(1:500; R&D Systems), goat anti-human CD26 (1:1000; R&D Systems), and mouse antibody to glyceraldehyde-3-phosphate dehydrogenase (1:4000; Santa Cruz).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Flow Cytometry, Derivative Assay, Polymerase Chain Reaction

Journal: Nature

Article Title: Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC.

doi: 10.1038/nature12005

Figure Lengend Snippet: Figure 4 | DPP4 is essential for virus infection. a, Inhibition of hCoV-EMC infection of Huh-7 cells by antibodies to DPP4. Supernatants collected at 2 h (open bars) and 20 h (filled bars) were tested for the presence of hCoV-EMC RNA using a TaqMan assay. Results representative of three different experiments are shown as DCt values (one-way ANOVA test, *P , 0.05; n 5 3 per group), normal goat, normal goat serum. b, Infection of human primary bronchiolar epithelial cells is blocked by the DPP4 antibodies in a dose- dependent manner and samples were analysed at 2 h (open bars) and 20 h (filled bars) after infection (one-way ANOVA test, *P , 0.05; n 5 3 per group). c, COS-7 cells transfected with plasmids encoding human DPP4 (hDPP4), bat DPP4 (bDPP4) or a control plasmid (pcDNA) were inoculated with hCoV-EMC at a multiplicity of infection of 1 and left for 1 h. Cells were washed twice and stained at 8 h after infection (original magnification, 3200) or supernatant collected at 2 h (open bars), 20 h (black bars) and 40 h (blue bars) was tested for the presence of hCoV-EMC RNA using a TaqMan assay (d). Results representative of three different experiments are expressed as GE (TCID50 ml21) values (one-way ANOVA test, *P , 0.05; n 5 4 per group). All error bars represent s.e.m.

Article Snippet: The cryosectionswere immunostained using the procedure as described26, using mouse monoclonal antibody anti-b-tubulin IV (Sigma) and goat anti-hDPP4 polyclonal antibody (R&D Systems) as primary antibodies, and Dylight-488-labelled, anti-mouse IgG (H1L) andDylight-549-labelled, anti-goat IgG (H1L) as secondary antibodies (Jackson ImmunoResearch).

Techniques: Virus, Infection, Inhibition, TaqMan Assay, Transfection, Control, Plasmid Preparation, Staining