Skip to main content
. Author manuscript; available in PMC: 2020 Mar 1.
Published in final edited form as: Cancer Immunol Res. 2019 Jul 30;7(9):1547–1561. doi: 10.1158/2326-6066.CIR-18-0367

Figure 2: Thrombospondin-1 inhibits NK cell proliferation and activation.

Figure 2:

NK cells were isolated from spleens of WT mice by MACS NK isolation kit. (A) Cells were cultured in serum free RPMI for 24 hours with/out thrombospondin-1 (2 nM) followed by fixation, permeabilization and i.c. staining for Ki-67, n = 3 biological replicates, error bars indicate standard error of the mean (SEM). (B) NK cells were cultured in serum free RPMI with/without IL-15 (10 ng/ml) and MTS absorbance was measured after 24 hrs of culture in the presence or absence of thrombospondin-1 (2 nM), n = 7 biological replicates, error bars indicate SEM. (C) Isolated NK cells were divided into 4 groups and cultured in serum free RPMI with thrombospondin-1 (2 nM), LPS (1 μg/ml) or both. mRNA abundance (relative to β-Actin) and (D) cell surface protein levels of CD69 were measured by RT-qPCR (LPS versus TSP-1+LPS treatments, P = 0.0373, n = 3 biological replicates, error bars indicate SEM) and flow cytometry (representative), respectively. (E) Human NK-92 cells were cultured in serum free RPMI with 100 IU IL-2 and treated either with mouse IgG (1 ug/ml) or B6H12 (1 ug/ml) in the presence or absence of human thrombospondin-1 (2 nM) and MTS absorbance was measured after 48 hrs, n = 5 technical replicates, error bars indicate SEM.