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: Oral Science International

Article Title: Down‐regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma

doi: 10.1016/s1348-8643(12)00004-3

Figure Lengend Snippet: Fig. 1. Family with sequence similarity 3, member B (FAM3B) mRNA expression in oral squamous cell carcinoma (OSCC)-derived cell lines. FAM3B mRNA levels were

Article Snippet: Immunohistochemical analysis Immunohistochemical analysis was performed on 4- m sections of paraffin-embedded specimens using anti-FAM3B polyclonal antibody (Proteintech).

Techniques: Sequencing, Expressing, Derivative Assay

Journal: Oral Science International

Article Title: Down‐regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma

doi: 10.1016/s1348-8643(12)00004-3

Figure Lengend Snippet: Fig. 2. Family with sequence similarity 3, member B (FAM3B) protein expression in oral

Article Snippet: Immunohistochemical analysis Immunohistochemical analysis was performed on 4- m sections of paraffin-embedded specimens using anti-FAM3B polyclonal antibody (Proteintech).

Techniques: Sequencing, Expressing

Journal: Oral Science International

Article Title: Down‐regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma

doi: 10.1016/s1348-8643(12)00004-3

Figure Lengend Snippet: Fig. 3. Expression profile of family with sequence similarity 3, member B (FAM3B) mRNA

Article Snippet: Immunohistochemical analysis Immunohistochemical analysis was performed on 4- m sections of paraffin-embedded specimens using anti-FAM3B polyclonal antibody (Proteintech).

Techniques: Expressing, Sequencing

Journal: Oral Science International

Article Title: Down‐regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma

doi: 10.1016/s1348-8643(12)00004-3

Figure Lengend Snippet: Fig. 4. Immunohistochemical (IHC) analysis of family with sequence similarity 3, member B (FAM3B) protein expression in primary oral squamous cell carcinoma (OSCC) samples. Representative results of IHC analysis for FAM3B in normal oral tissue (A, B) and primary OSCC (C, D). Original magnification: ×100 (A, C) and ×400 (B, D). Positive immunoreactions

Article Snippet: Immunohistochemical analysis Immunohistochemical analysis was performed on 4- m sections of paraffin-embedded specimens using anti-FAM3B polyclonal antibody (Proteintech).

Techniques: Immunohistochemical staining, Sequencing, Expressing