Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: Sl-1 cells infected with AfMNPV for 6 h were co-stained with anti -cytochrome c monoclonal antibody and Mito-Tracker red. After incubation with FITC-coupled secondary antibodies, cells were visualized by confocal laser scanning microscopy. Con: Control cells treated without AfMNPV; Inf: Cells infected with AfMNPV for 6 h.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Infection, Staining, Incubation, Confocal Laser Scanning Microscopy

Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: (A) Identification of recombinant plasmid pLitmus-cytc containing cytochrome c DNA fragment. pLitmus-cytc was digested with BamH I and EcoR I and then analyzed by agarose gel electrophoresis. M1: 1 kb maker, M2: 100 bp maker, S: plasmid DNA products. (B) Electrophoretic analysis of dsRNA transcribed in vitro . pLitmus-cyt c was digested with BamH I and EcoR I respectively and transcribed in vitro , and RNA products were electrophorised on agrose gel. M: 100 bp maker, S: RNA transcription products. (C) Cytochrome c mRNA level after treatment with dsRNA. Sl-1 cells treated with dsRNA for 0, 24, 48, and 72 h, and after inoculated with AfMNPV for 10 h, total RNA in each treatment was isolated. Cytochrome c mRNA was determined by semi-quantitative RT-PCR.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Recombinant, Plasmid Preparation, Agarose Gel Electrophoresis, In Vitro, Isolation, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: (A) Sl-1 cells transfected with GFP dsRNA; (B) Sl-1 cells transfected with cyt-c dsRNA. M, protein marker. β-tubulin was used as an internal control. *: non-specific band.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Transfection, Marker

Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: (A) Microscopy image of cells treated with AfMNPV and cyt-c dsRNA. 1. Cells infected with AfMNPV alone for 10 h; 2. Cells infected with AfMNPV for 10 h after treatment with cyt c dsRNA for 24 h; 3. Cells infected with AfMNPV for 10 h after treatment with cyt-c dsRNA for 48 h.; 4. Cells infected with AfMNPV for 10 h after treatment with cyt-c dsRNA for 72 h. (B) Flow cytometric analysis. Sl-1 cells treated with dsRNA for 48 h,and infected with AfMNPV. At 10 h post infection, cells were staining by PI and FITC-Annexin. Apoptosis was analyzed by flow cytometry. (C) Cyt-c dsRNA resulted in the decrease of apoptosis induced by AfMNPV in Sl-1cells. Data were representive for three independent experiments. *, p <0.05.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Microscopy, Infection, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: (A) Sl-1 cells treated with dsRNA for different time points, apoptosis was induced with AfMNPV, and then caspase-3 activity was measured at 10 h post-infection.1. Control cells treated alone with AfMNPV; 2. Cells treated with GFP dsRNA and virus; 3. Cells without any treatment; 4. Cells treated with dsRNA, 24 h later, infected by AfMNPV for 10 h; 5. Cells treated alone with dsRNA for 24 h; 6. Cells treated with dsRNA, 48 h later, infected by AfMNPV for 10 h; 7. Cells treated alone with dsRNA for 48 h; 8. Cells treated with dsRNA, 72 h later, infected by AfMNPV for 10 h; 9. Cells treated alone with dsRNA for 72 h. (B) Caspase-9 activity in cytochrome c dsRNA-treated Sl-1 cells after infection with AfMNPV for 10 h, compared with control cells.1. Normal cells; 2. Cells infected with AfMNPV; 3. Cells infected with AfMNPV for 10 h after GFP dsRNA treatment for 48 h; 4. Cells infected with AfMNPV for 10 h after cyt c dsRNA treatment for 48 hr. *, p <0.05.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Activity Assay, Infection

Journal: PLoS ONE

Article Title: The Role of Cytochrome c on Apoptosis Induced by Anagrapha falcifera Multiple Nuclear Polyhedrosis Virus in Insect Spodoptera litura Cells

doi: 10.1371/journal.pone.0040877

Figure Lengend Snippet: (A) The treatment of FSBA resulted in the decrease of the percentage of apoptotic cells under the inductuion of AfMNPV. (B) The treatment of FSBA inhibited activation of pro-caspase-3 in AfMNPV-infected Sl-1 cells. *, There were significant differences.

Article Snippet: SL-1 cells were propagated in GIBCO™ Grace's medium (Invitrogen, Grand Island, N.Y., USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, N.Y., USA), 0.3% yeast extract and 0.3% lactalbumin hydrolysate at 28°C.

Techniques: Activation Assay, Infection