Figure 6: H60b mediates increased cytokine expression of NK Cells by Sirt6 heterozygosity. | Scientific Reports

Figure 6: H60b mediates increased cytokine expression of NK Cells by Sirt6 heterozygosity.

From: Epigenetic regulation of NKG2D ligands is involved in exacerbated atherosclerosis development in Sirt6 heterozygous mice

Figure 6

(A) Schematic diagram of the experiments. Peritoneal macrophage, isolated from ApoE−/− and Sirt6+/−ApoE−/− mice, were incubated to adhere to culture plate. Cells were treated with 30 μg/ml ox-LDL for 24 hours, and cocultured with NK cells, at 1:10 ratio for 6 hours. The NK cells were collected and used for determination of cytokine expression by realtime PCR. (B,C) Macrophages were transfected with Si-Ctrl or Si-H60b to knockdown H60b, cultured for 24 hours, treated with 30 μg/ml ox-LDL for 24 hours, and then co-incubated with NK cells at 1:10 ratio for 6 hours. (B) Macrophages were collected to determinate Sirt6 and H60b expression at the mRNA level and protein level. (C) The NK cells were collected and used for determination of cytokine expression by realtime PCR. (D) To block ligand-receptor interaction, isotype IgG antibody or NKG2D antibody (R&D, BAM1547, 3 μg/ml) or H60b antibody (R&D, MAB1155, 3 μg/ml) were added for 2 hours before co-incubation with NK cells. The NK cells were collected and used for determination of cytokine expression by realtime PCR. P value was obtained by two-way analysis of variance (ANOVA) plus a post hoc analysis using the Bonferroni test. (*P < 0.05, **P < 0.01, ***P < 0.005).

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