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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: Phytosomal curcumin causes natural killer cell-dependent repolarization of glioblastoma (GBM) tumor-associated microglia/macrophages and elimination of GBM and GBM stem cells

Fig. 2

CCP treatment is associated with intra-tumor recruitment of activated NK cells, which is eliminated in mice peripherally injected with the NK cell-neutralizing NK1.1 antibody (NK1.1Ab). a After intracranial implantation of 105 GL261 cells on day 1, the GBM-mice (n = 17) were randomly divided into four groups: ‘Vehicle’ (n = 5), ‘CCP’ (n = 5), ‘CCP + NK1.1’ (n = 4), and ‘NK1.1’ (n = 3). On day 11, the CCP + NK1.1 and NK1.1 groups received the NK1.1Ab (100 μg/mouse), whereas the other two groups received mouse IgG (as a negative control) (100 μg/mouse) (i.p.).On day 12, each mouse received either CCP (2 mg/ mouse/day in 200 μl PBS) for the ‘CCP’ and ‘CCP + NK1.1’ groups or PBS for the ‘Vehicle’ and ‘NK1.1’ groups for five days. On day 17, all mice were sacrificed and the GBM tumor in each mouse was cut into two parts and processed for IHC (fixed and sectioned) and flow cytometry (dispersed by trypsinization and fixed), respectively. b and c IHC: Four randomly chosen GBM-containing brain sections per mouse from each group were stained with the NKp46 antibody. The Vehicle-treated mice displayed very weak and sparse NKp46 staining (b top row and c), but the CCP-treated showed a 200% increase in NKp46 fluorescence (b middle row and c). This CCP-evoked NKp46 staining was virtually eliminated in the NK1.1Ab-treated mice (b bottom row and c) (*p = 1.2 × 10− 8 CCP versus Vehicle-treated; ∆ p = 7.3 × 10− 9 CCP versus CCP + NK1.1; **p = 1.0 × 10− 3 CCP + NK1.1 versus Vehicle-treated) (Scale bar: 47.62 μm). d-f Flow cytometry: CCP-treated samples displayed a 190% increase in integrated NKp46 fluorescence (IF) (*p = 0.02, CCP versus Vehicle). The NKp46 IF was virtually eliminated in the CCP + NK1.1 samples (f, ∆ p = 0.04, CCP + NK1.1 versus CCP; **p = 5.0 × 10− 3, CCP + NK1.1 versus Vehicle-treated). Data (mean ± S.D.) obtained from Vehicle (n = 4), CCP (n = 4), and CCP + NK1.1 (n = 4). Integrated fluorescence (IF) = mean fluorescence per cell X total number of cells (events) in a segregated population. d, e The NKp46(+) events (NK cells) were virtually eliminated in the NK1.1 samples

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