27,28. During cell-to-cell interactions, it acts both as an adhesion and signaling molecule29. Following its activation, SLAMF5 recruits adaptors of the SLAM adaptor protein (SAP) family which induces downstream signaling. SLAMF5 was previously reported to mediate the interactions...
Where indicated, CellTrace Violet (Thermofisher Scientific) labeling of cells was conducted as per manufacturer’s instructions, used at a final concentration of 2 μM prior to stimulation. All inhibitors and metabolites were extensively titrated prior to use, and reagent details are as follows:...
Total RNA was extracted from BMDMs or bone marrow using TRIzol reagent (21101, AG) according to the manufacturer’s instructions. cDNA was synthesized using a reverse transcription kit (11706, AG), and a SYBR green real-time PCR premix kit (11701, AG) was used to quantify the expression of...
Cells were then fixed and stained according to instructions by the manufacturer, using an intracellular cytokine staining kit (eBioscience, San Diego, CA). Following antibodies were used for staining: Thy1.2− APC/PE-Cy7/FITC (allophycocyanin/ phycoerythrin-Cy7/fluorescein isothiocyanate), Thy1.1...
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/mL; Sigma-Aldrich), ionomycin (250 ng/mL; Sigma-Aldrich) and brefeldin A (BD bioscience, San Jose, CA, USA) were added for the last 4 h of cell culture, and cells were fixed and permeabilized using a Cytofix/Cytoperm kit (BD Biosciences) according to the manufacturer’s instructions....
2009 W-2 Form InstructionsStatement, Reissued
(14,000×g, 10 min) and endogenous ubiquitylated proteins were isolated from the soluble lysate at 4 °C for 3–20 h using 20 µl of packed agarose TUBE (TUBE1, Life sensors, performed according to the manufacturer’s instructions) or 20 µl of GST-UBA beads (generated ...
IL-22 levels in cell culture supernatant were measured using a mouse IL-22 ELISA kit (Antigenix America) according to the manufacturer’s instructions. TGFβ abundance in lysates from frozen human tumor and control tissue was determined by TGFβ1 Emax ImmunoAssay System (Promega) according to th...
Cells were washed three times with pre-warmed sterile PBS, and total RNA was extracted from the cells in 1 mL of TRIzol reagent (Ambion) according to the manufacturer’s instructions. Extracted RNA was treated with DNase I (Roche) and precipitated with sodium acetate (Ambion) and ethanol. ...