il-10 inhibitor as101  (Tocris)

Journal: Juntendo Medical Journal

Article Title: The Role of Tumor-associated Macrophages in Colorectal Peritoneal Metastasis in Mice

doi: 10.14789/ejmj.JMJ24-0049-OA

Figure Lengend Snippet: (A) Protocol for macrophage inhibitor (clodronate) and an IL-10 inhibitor (AS101) injection into NOD SCID mice. (B) Macroscopic views and the mean weight of the peritoneal dissemination nodules from animals injected with clodronate and negative controls. The administration of clodronate significantly reduced the size of peritoneally disseminated nodules (P < 0.001). (C) Macroscopic views and the mean weight of the peritoneally disseminated nodules from animals injected with AS101 and negative controls.

Article Snippet: An IL-10 inhibitor AS101 (Tocris Bioscience, Bristol, UK) was injected (20 mg/mouse) on days 1-35 (control group: 4 mice, AS101 group: 3 mice).

Techniques: Injection

Journal: PLoS Pathogens

Article Title: IL-10-mediated signals act as a switch for lymphoproliferation in Human T-cell leukemia virus type-1 infection by activating the STAT3 and IRF4 pathways

doi: 10.1371/journal.ppat.1006597

Figure Lengend Snippet: A . ILT cells were transfected with control siRNA (si-CTRL) (open bar) or si-STAT3 (black bar), and subjected to qRT-PCR 48 h after electroporation. Indicated gene expression levels were measured and standardized with ACTB ( β-actin ) mRNA levels in each sample. The relative values against the si-CTRL samples are presented as means and SD of duplicate samples. Representative results of two independent experiments are shown. * p <0.05, ** p <0.01. B. ILT-22 and ILT-294 cells cultured with (■) or without (□) rhIL-10 for at least 1 week, and IL10 mRNA levels were measured. The normalized values against ACTB were indicated as means and SD of duplicate samples. N.D., not detected. C. ILT-H2 (●), ILT-22 (▲), and Jurkat (◯) cells were incubated with indicated concentrations of AS101 or vehicle control (0.13% ethanol) for 5 days and the viable cell number was assessed using Cell Counting Kit-8. Relative values against vehicle control were plotted as means and SD of duplicate samples. D . ILT-H2 (▲), ILT-22 (●), ILT-294 (+) (cultured with IL-10) (■), and ILT-294 (-) (cultured without IL-10) (□) cells were incubated with indicated concentrations of Cucurbitacin I or vehicle control (0.1% DMSO) for 2 days, and the cell number was evaluated by Cell Counting Kit-8. The relative values against vehicle controls indicate means and SD of duplicate samples. E. Cell lysates of ILTs were harvested 48 h (ILT-294, ILT-22) or 72 h (ILT-H2) after transfection with si-CTRL or si-STAT3, probed with antibodies to caspase-3, cleaved caspase-3, survivin, and IRF4 in immunoblotting assay, and presented together with the β-actin immunoblots of corresponding membranes. Approximate sizes of caspases are indicated. a , b , denote the same images for β-actin because the same membranes were used for the detection of caspases and IRF4, respectively. ILT-294 cells were cultured in the presence of IL-10 in A and E .

Article Snippet: AS101, the non-toxic tellurium IL-10-inhibitor (Tocris, Ellisville, MO) [ ] and cucurbitacin I (JSI-124), a STAT3-inhibitor (Tocris) [ ] were also used.

Techniques: Transfection, Control, Quantitative RT-PCR, Electroporation, Gene Expression, Cell Culture, Incubation, Cell Counting, Western Blot