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. 2022 Jul 11;12(12):5451–5469. doi: 10.7150/thno.72248

Figure 2.

Figure 2

Knockdown of NONO suppresses proliferation and invasion, and promotes apoptosis in GBM cell lines. A CCK-8 assay for cell viability of NHA, NHA-ET, U251 and P3 after transfection with siNC or two different siRNAs (siNONO-1 and siNONO-2) (n = 3). Obtained data were normalized to the siNC group. B EdU assay to assess the cell growth of U251 transfected with siNC, siNONO-1 and siNONO-2 (n = 3). Scale bar = 100 μm. C 3D tumor spheroid invasion assay to measure invasion of U251 transfected with siNC, siNONO-1 and siNONO-2 (n = 3). Scale bar = 200 μm. D Model and representative images of co-culture invasion assays for U251 infected with lentiviral constructs expressing shNC or shNONO (n = 3). The invasion ability was evaluated at 72 h. Scale bar = 200 μm and 100 μm (magnified inset). E Flow cytometry analysis of Annexin V-FITC and propidium iodide (PI) staining for the detection of apoptosis in U251 and P3 transfected with siNC, siNONO-1 and siNONO-2 (n = 3). F Bioluminescence images of mice (n = 5 per group) orthotopically implanted with luciferase expressing U251 and P3 cells infected with lentiviral constructs expressing shNC or shNONO. Bioluminescence was collected to assess tumor growth. G Kaplan-Meier survival curve of groups of xenograft bearing mice (n = 5 per group). The Log-rank test was used to obtain statistical significance. H IHC for NONO and Ki67 levels in GBM xenografts. Scale bar = 100 μm. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.