Journal: Nature Communications

Article Title: A non-canonical pathway regulates ER stress signaling and blocks ER stress-induced apoptosis and heart failure

doi: 10.1038/s41467-017-00171-w

Figure Lengend Snippet: Mechanism for AGGF1 regulation of miR-183-5p expression. a ChIP–qPCR analysis for the interaction between ZEB1 and miR-183-5p promoter DNA ( n = 4/group, ** P < 0.01). b Schematic diagram showing that the miR-183-5p promoter-luciferase reporter with two ZEB1-binding motifs. c Luciferase activity ( n = 6/group, ** P < 0.01). d Real-time RT-PCR analysis for miR-183-5p expression in H9C2 cells after AGGF1 treatment with or without ZEB1 overexpression ( n = 3/group, ** P < 0.01). e Western blot analysis for the ZEB1 level in H9C2 cells after AGGF1 treatment ( n = 5/group, * P < 0.05). f Western blot analysis for activation of ERK1/2 in H9C2 cells after AGGF1 treatment ( n = 5/group, ** P < 0.01). g Real-time RT-PCR analysis for ERK1 and ERK2 expression by ERK siRNA ( n = 3/group, ** P < 0.01). h Western blot analysis for the ZEB1 level by ERK siRNA ( n = 5/group, * P < 0.05). i Schematic model for a non-canonical ER stress signaling pathway mediated by AGGF1. Data are shown as the mean ± s.d. from at least three independent experiments. Statistical analysis was carried out by a Student’s two-tailed t -test

Article Snippet: Protein–DNA complexes were immunoprecipitated with IgG or an anti-ZEB1 antibody (2 μg; Proteintech).

Techniques: Expressing, ChIP-qPCR, Luciferase, Binding Assay, Activity Assay, Quantitative RT-PCR, Over Expression, Western Blot, Activation Assay, Two Tailed Test

Journal: Scientific Reports

Article Title: A canine chimeric monoclonal antibody targeting PD-L1 and its clinical efficacy in canine oral malignant melanoma or undifferentiated sarcoma

doi: 10.1038/s41598-017-09444-2

Figure Lengend Snippet: Enhancement of cytokine production and cell proliferation of dog peripheral blood mononuclear cells by c4G12 treatment. Dog peripheral blood mononuclear cells ( n = 7) were obtained from healthy beagle donors and stimulated by 5 μg/mL staphylococcal enterotoxin B in the presence or absence of 20 μg/mL c4G12. Dog IgG was used as a control antibody. For evaluation of cytokine production, the culture supernatant was harvested on day 3, and concentration of ( a ) IL-2 or ( b ) IFN-γ was measured by ELISA. To evaluate cell proliferation, nucleotide analogue 5-ethynyl-2′-deoxyuridine (EdU) was added to the medium on day 2, and cells were harvested after incubation for another 2 h. The lymphocyte population was gated by forward scatter and side scatter, and the incorporation of EdU in ( c ) CD4+ or ( d ) CD8+ cells was measured by a flow cytometer. Statistical analysis was performed with a Wilcoxon signed rank-sum test.

Article Snippet: For cytokine assays, the culture supernatant was harvested on day 3 and the concentrations of IL-2 and IFN-γ were measured by Duoset ELISA canine IL-2 and IFN-γ (R&D systems, Minneapolis, MN, USA), respectively, according to the manufacturer’s instructions.

Techniques: Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Flow Cytometry