Figure 8 | Scientific Reports

Figure 8

From: Dual-sgRNA CRISPR/Cas9 knockout of PD-L1 in human U87 glioblastoma tumor cells inhibits proliferation, invasion, and tumor-associated macrophage polarization

Figure 8

Functional polarization of TAMs by PD-L1 knockout in U87 cells. U87 cells were treated with Cas9-g82/165 + HDR for 24 h, then human macrophages were added allowing TAM differentiation for 48 h. An ELISA assay was used to detect (A) human IL-4 secretions in the medium from co-cultured human macrophages and U87 cells with or without Cas9-g82/165 + HDR. In parallel, mouse BMMs co-cultured with U87 cells were investigated to examine PD-L1 knockout on mouse (B) TNF-α and (C) IL-4 secretions. PD-L1 deletion in U87 cells significantly upregulated TNF-α levels and reduced IL-4 secretion compared to the control. All experiments were performed in triplicate and results were normalized for direct comparisons. Data represents the mean ± SEM. Statistical significance was evaluated with a t-test. *p < 0.05, **p < 0.01.

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