Journal: Journal of Integrative Agriculture

Article Title: Critical role of cytochrome c1 and its cleavage in porcine reproductive and respiratory syndrome virus nonstructural protein 4-induced cell apoptosis via interaction with nsp4

doi: 10.1016/s2095-3119(17)61670-8

Figure Lengend Snippet: Fig. 1 Porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 4 (nsp4) interacts with cytochrome c1 (cyto.c1). A, yeast two hybrid assay. The plasmids pGBKT7-nsp4 and pGADT7-cyto.c1 were co-transformed into the yeast strain Y2HGold and selected on QDO/X/ABA plates. The top and second panels are the respective positive and negative controls. B, HEK 293T cells were transfected to express Flag-cyto.c1, or HA-nsp4 or together. At 24 h post-transfection, the cells were either subjected to direct Western blot analysis or lysed and immunoprecipitated with anti-HA antibodies. The proteins bound to sepharose beads were separated by SDS-PAGE, transferred to PVDF membranes, and probed with the antibodies to Flag and HA. C, the same as B except that nsp4 truncation mutants were used. D, MARC-145 cells were transduced with letiviruses that were expressing GFP or nsp4-GFP in the presence of 8 µg mL–1 polybrene, respectively. The cells were harvested 48 h post-transduction and cell lysates were immunoprecipitated using anti-GFP beads. The proteins bound to sepharose beads were subjected to Western blot analysis using an anti-cytochrome c1 polyclonal antibody or anti-GFP mAb. E, PRRSV nsp4 interacts with cyto.c1 in mammalian cells. Vero cells grown on coverslips in six-well plates were transfected when 60–70% to express the indicated proteins, either alone or pairwise. At 18–24 h post-transfection, the cells were fixed, permeablized, and stained with antibodies to HA and Flag, and examined by confocal microscopy. For double transfections, the merged images are shown at the right.

Article Snippet: Mouse anti-green fluorescent protein (GFP) mAb (66002-1-Ig) and rabbit anti-cytochrome c1 polyclonal antibodies (10242-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: Virus, Y2H Assay, Transformation Assay, Transfection, Western Blot, Immunoprecipitation, SDS Page, Transduction, Expressing, Staining, Confocal Microscopy

Journal: Urology

Article Title: Preliminary immunohistochemical characterization of a monoclonal antibody (pro:4-216) prepared from human prostate cancer nuclear matrix proteins

doi: 10.1016/s0090-4295(97)00337-3

Figure Lengend Snippet: FIGURE 2. NMP composition of normal human prostate (A) and human prostate cancer (C) taken from a radical prostatectomy specimen from a patient with clinical/y localized prostate cancer. High resolution two-dimensional gel electrophoresis of NMP preparations were isolated and analyzed as described in Methods. Panels B and D represent Western immunoblots using PRO:4-2 16 monoclonal antibody; immunoblotting technique as described in Methods. Arrowhead represents the localization of NMP(s) with immunoreactivity to the PRO:4-2 16 mouse monoclonal anti- body generated against human prostate cancer nuclear matrix preparations. kD = molecular weight in thousands; SDS-PAGE = sodium dodecyl sulfate-polyacrylamide gel electrophoresis; pl = isoelectric point.

Article Snippet: The mouse monoclonal antibodies were UROLOGY 50 (51, 1997 801 then detected with a CY3 conjugated goat anti-mouse antibody (Jackson ImmunoResearch, West Grove, Pa) and DNA was visualized with DAPI (4,6-diamidino-2-phenylindole) at 2 /.

Techniques: Two-Dimensional Gel Electrophoresis, Electrophoresis, Isolation, Western Blot, Generated, Molecular Weight, SDS Page, Polyacrylamide Gel Electrophoresis

Journal: Urology

Article Title: Preliminary immunohistochemical characterization of a monoclonal antibody (pro:4-216) prepared from human prostate cancer nuclear matrix proteins

doi: 10.1016/s0090-4295(97)00337-3

Figure Lengend Snippet: FIGURE 3. lmmunohistochemical staining of human prostate tissue with the mouse monoclonal antibody PRO:4- 2 16. Anti-mouse IgM secondary antibody (Vector’s ABC Elite Kit). 3-3’-Diaminobenzidine tetrahydrochloride (DAB) was used as a chromogen following ethyl green counterstain. (A) Histologically normal prostate tissue (nuclei dem- onstrate an IHC intensity score of 0). (B) Histologically normal prostate tissue with the emphasis on stromal nuclei (weakly positively staining stromal nuclei represent an IHC intensity score of 1). (Cl Adenocarcinoma of the prostate from a tumor with Gleason grade 3 + 3 = 6 (IHC intensity score of 3). The cancerous glands surround a noncancerous gland. (0) A prostatic gland demonstrating PIN (positively staining nuclei represent an IHC intensity score of 1 to 2). The arrow in panel B identifies an area of prostatic stroma. The small arrows in panel C identify prostatic ade- nocarcinoma glands and the large arrow in panel C identifies a histologically normal gland found among cancerous glands. The arrow in panel D represents histologic PIN.

Article Snippet: The mouse monoclonal antibodies were UROLOGY 50 (51, 1997 801 then detected with a CY3 conjugated goat anti-mouse antibody (Jackson ImmunoResearch, West Grove, Pa) and DNA was visualized with DAPI (4,6-diamidino-2-phenylindole) at 2 /.

Techniques: Staining

Journal: British Journal of Nutrition

Article Title: Mitochondrial dysfunction in the liver of type 2 diabetic Goto–Kakizaki rats: improvement by a combination of nutrients

doi: 10.1017/s0007114511000493

Figure Lengend Snippet: Fig. 5. Apoptosis-related factors p53 and p21 in liver tissue: (a) Western blotting images of p21, p53 and a-tubulin, respectively and (b) quantitative results of p53 and p21. Quantitative values were computed as the ratios of the density of the targeted protein to a-tubulin. Results are expressed as fold change of control. Values are means, with their standard errors represented by vertical bars (n 10). Mean value was significantly different from that of the Wistar group: * P,0·05, ** P,0·01. , Wistar; , GK; , GK with nutrients; , GK with pioglitazone.

Article Snippet: Liver tissue proteins were subjected to 10 % SDS-PAGE and detected with primary antibodies against p53 (1:1000, Mouse Ab no. 12 506; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p21 (1:1000, Mouse Ab no. 2946; Cell Signaling Technology, Danvers, MA, USA) or a-tubulin (1:5000).

Techniques: Western Blot, Control