Journal: Cancer cell

Article Title: A Potent and Selective Small-molecule Degrader of STAT3 Achieves Complete Tumor Regression in vivo

doi: 10.1016/j.ccell.2019.10.002

Figure Lengend Snippet: SD-36-mediated Cellular STAT3 Protein Degradation (A) Cells were treated with SD-336, 10 μM of SI-109 or SD-36Me for 3 hr (MOLM-16) or 24 hr (SU-DHL-1) for immunoblotting. (B) MOLM-16 cells were treated with 0.25 μM of SD-36 for immunoblotting. (C) MOLM-16 cells stably transduced with vector control or CRBN shRNAs were treated with 1 μM of SD-36 for 3 hr for immunoblotting. (D) MDA-MB-231 cells transfected with the indicated STAT3-expressing vectors were treated with 1 μM of SD-36 for 16 hr for immunoblotting. (E) Human PBMCs were treated with SD-36 or 10 μM of SD-36Me for 24 hr for immunoblotting. (F) Cells were treated with 10 μM of SD-36 for 2.5 hr for multiplexed quantitative proteomics analysis. p value: Student’s t-test. (G) Cells were treated with 1 μM of the indicated compounds for 24 hr for immunoblotting. (H) Parental DLD-1 (WT) and DLD-1/STAT3 Y705F/Y705F (Y705F) cells were pre-incubated with or without 10 ng/ml of IL-6 for 15 min, then 1 μM of SD-36 for 24 hr for immunoblotting. .

Article Snippet: Cell Lines MDA-MB-231, A20 and human PBMCs (PCS-800–011) were obtained from ATCC.

Techniques: Stable Transfection, Transduction, Plasmid Preparation, Multiple Displacement Amplification, Transfection, Expressing, Incubation

Journal: BMC Veterinary Research

Article Title: Evaluation of the limitations and methods to improve rapid phage-based detection of viable Mycobacterium avium subsp. paratuberculosis in the blood of experimentally infected cattle

doi: 10.1186/s12917-016-0728-2

Figure Lengend Snippet: Reproducibility of detecting MAP from whole blood using PMMS and from PBMCs. Figure show the reproducibility of detecting viable MAP cells using paired samples tested independently with the phage assay using the PMMS to isolate the MAP cells from whole blood ( a ) or detecting the MAP cells with the phage assays directly from PBMCs isolated from sheep blood ( b )

Article Snippet: Bacterial strains, bacteriophage and growth medium To optimise MAP detection in the peripheral blood mononuclear cells (PBMCs), MAP strains K10 and ATCC 19698 were used.

Techniques: Isolation

Journal: Frontiers in Oncology

Article Title: A Potent and Selective Dual Inhibitor of AXL and MERTK Possesses Both Immunomodulatory and Tumor-Targeted Activity

doi: 10.3389/fonc.2020.598477

Figure Lengend Snippet: Inhibition of phagocytosis and reversal of macrophage-mediated suppression of T cells by INCB081776 in primary human immune cells. (A) Human whole blood was treated with INCB081776 at 10 nM and 100 nM along with dimethyl sulfoxide control. Fluorescent microspheres were added to peripheral blood mononuclear cells (PBMCs) prelabeled with anti-CD14 and anti-CD16, and acquisition performed at intervals over a course of 4 min. Plot indicating the mean of the uptake of fluorescent microspheres by CD14 ++ CD16 + monocyte population over time is shown. Data are representative of three healthy donors. (B) Primary macrophages differentiated in vitro from human PBMCs were pretreated for 2 h with INCB081776 followed by stimulation with the MERTK-specific agonist antibody MAB8912 for an additional 30 min. Levels of phospho-MERTK (pMERTK) and total MERTK were determined by Western blot. (C) Primary macrophages differentiated in vitro from human PBMCs were pretreated with various concentrations of INCB081776 overnight, followed by incubation with carboxyfluorescein succinimidyl ester (CFSE)-labeled T cells for an additional 5 days. T-cell proliferation was determined by FACS analysis ( N = 2 per group). (D) Following incubation of macrophages and T cells in the presence of increasing concentrations of INCB081776 as in panel (C) , supernatants were collected and interferon (IFN)-γ cytokine production was measured ( N = 2 per group, except for 111 and 333 nM [ N = 1]). SD, standard deviation.

Article Snippet: Human peripheral blood mononuclear cells (PBMCs) were obtained from normal leukapheresis of two healthy donors (Biological Specialties, Colmar, PA, USA).

Techniques: Inhibition, In Vitro, Western Blot, Incubation, Labeling, FACS, Standard Deviation