Additional file 1 of Inhibitory receptor CD47 binding to plasma TSP1 suppresses NK-cell IFN-γ production via activating the JAK/STAT3 pathway during HIV infection
Creators: name: Lang, Bin
name: Wang, Meiting
name: Zhang, Zining
name: Fu, Yajing
name: Han, Xiaoxu
name: Hu, Qinghai
name: Ding, Haibo
name: Shang, Hong
name: Jiang, Yongjun
Publisher: name: figshare
Year: 2023
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Figure S1. TSP1–CD47 axis inhibits NK cells activation, proliferation, and IFN-γ production. A Schematic diagram of experiments and representative cytometry dot plots using NK cells from NC group treated with different concentrations of recombinant TSP1 (0, 250, 500, and 1000 ng/mL) to study TSP1–CD47 axis effects on proliferation (Ki-67 expression) and activation (CD69 expression) of human NK cells. B Schematic diagram of experiments and representative cytometry dot plots using NK cells from NC group treated with recombinant TSP1 (1000 ng/mL) or Anti-CD47 antibody (2 μg/mL) to study TSP1–CD47 axis effects on IFN-γ production of human NK cells. C Paired comparisons of the percentage of IFN-γ expression in total NK cells with recombinant TSP1 (1000 ng/mL) treatment and anti-CD47 antibody (2 μg/mL) or IgG-control (2 μg/mL) treatment (n = 7). Paired-t or Wilcoxon signed-rank test was used to make paired-group comparisons, and error bars represent median and interquartile range; *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001; ns, no significance; NT, no treatment. Figure S2. Blocking SIRPα restores IFN-γ production of NK cells via NFAT signaling pathway in PLWH. A Schematic diagram of experiments and representative cytometry dot plots using NK cells from people living with HIV treated with anti-SIRPα antibody (2 μg/mL) or IgG-control (2 μg/mL) to study effects on IFN-γ production of human NK cells. B Paired comparisons of the percentage of IFN-γ expression in total NK cells from the people living with HIV with anti-SIRPα antibody (2 μg/mL) or IgG-control (2 μg/mL) treatment (n = 7). C Schematic diagram of experiments and representative cytometry dot plots using NK cells from people living with HIV treated with Anti-SIRPα antibody (2 μg/mL) or IgG-control (2 μg/mL) to study effects on the phosphorylation of the nuclear factor of activated T-cells (NFAT) signaling pathway in human NK cells. Blue, Anti-SIRPα antibody treatment; red, IgG-control treatment. D Paired comparisons of the percentage of NFAT phosphorylation in total NK cells from people living with HIV with anti-SIRPα antibody (2 μg/mL) or IgG-control (2 μg/mL) treatment (n = 3). A paired t-test was used for paired-group comparisons, and error bars represent the median and interquartile range; **p &lt; 0.01, ***p &lt; 0.001. Figure S3. TSP1–CD47 axis activates STAT3 signal pathway of NK cells. A Schematic diagram of experiments and representative cytometry dot plots using NK cells from NC group treated with or without recombinant TSP1 (1000 ng/mL) to study TSP1–CD47 axis effects on phosphorylation of STAT3 signaling pathway of human NK cells. B Schematic diagram of experiments and representative cytometry dot plots using NK cells from people living with HIV treated with recombinant TSP1 (1000 ng/mL) and with or without STAT3-inhibitor Stattic (2 µmol/L) to study STAT3 blockade effects on IFN-γ production of human NK cells. Table S1. Characteristics of subjects enrolled in this study. Table S2. Key resources.