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. 2022 Aug 29;61(4):122. doi: 10.3892/ijo.2022.5412

Figure 2.

Figure 2

Effect of VEGF-A signaling in association with the expression patterns of NRP-1, on GB cells. (A and B) Nine GB cells were incubated in the absence or presence of exogenous VEGF-A (50 µg/l, left) or SU1498 (50 µmole/l, right), a VEGF receptor inhibitor, for 72 h, after which they were examined for cell viability using WST-1 assay and cell death using LDH assay (n=3; Tukey's post hoc test was applied to significant group effects in ANOVA, P<0.0001; asterisks indicate significant differences compared with the control group, *P<0.05 and **P<0.01). (C) Efficacy of Flt-1, Flk-1, NRP-1, and VEGF-A siRNA transfections assessed by western blot analysis. GAPDH was used as a control. (D) Cell death was assessed 72 h after transient transfection with siRNAs against VEGF-A, Flt-1, Flk-1, and NRP-1 using an LDH assay (n=3; Tukey's post hoc test was applied to significant group effects in ANOVA, P<0.0001; asterisks indicate significant differences compared to 0% inhibition; *P<0.05 and **P<0.01). (E) Effect of SU1498 (10 mg/kg) on the tumor volumes of U251-MG xenograft models (control group, n=4; SU1498 10 mg/kg, n=4) measured for 42 days using the following formula: V=0.523 LW2 (L=length and W=width). Bold arrows indicate the time-points of SU1498 injection (Tukey's post-hoc test was used to determine significant group effects in ANOVA, P<0.0001; asterisks indicate significant differences between the control group and the SU1498-injected group; *P<0.05 and **P<0.01). (F) Effect of SU1498 (10 mg/kg) on the tumor volumes of LN215-MG xenograft models (control group, n=4; SU1498 10 mg/kg, n=4) measured for 42 days using the formula aforementioned, V=0.523 LW2. Bold arrows indicate the time-points of SU1498 injection. VEGF-A, vascular endothelial growth factor-A; GB, glioblastoma; NRP-1, neuropilin-1; LDH, lactate dehydrogenase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; Flt-1, FMS related receptor tyrosine kinase 1; Flk-1, fetal liver kinase 1; n.s., non-significant; si-RNA or si-, small interfering RNA.