Figure 5 | Scientific Reports

Figure 5

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

Figure 5

PD-L1 knockout slows U87 tumor cell proliferation and growth. (A) BrdU staining was performed to analyze cell proliferation. Images suggest that U87 cells treated with Cas9-g82/165 + HDR proliferate at a slower rate compared to control cells. (B) Quantification of A. (C) Quantification of the growth rate of U87 cells transfected with or without Cas9-g82/165 + HDR. Cells were counted over 3 days. Images revealed that PD-L1 deletion inhibited U87 cell growth compared to the control. (D) An MTT assay was performed to examine U87 cell viability after treatment with Cas9-g82/165 + HDR for 2 days. Untreated cells (Ctl-U) and cells treated with lipofectamine only (Ctl-T) served as the controls. U87 cells treated with Cas9-g82/165 and Cas9-g82/165 + HDR displayed greater toxicity compared to the control and cells treated with the individual sgRNAs. All experiments were performed in triplicate and results were normalized for direct comparisons. The data represent the mean ± SEM, and statistical significance was evaluated with either a t-test (B and C) or a one-way ANOVA followed by Tukey’s post-hoc analysis (D). **p < 0.01. Scale bar = 100 µm.

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