Fig 1: Kaplan-Meier survival analysis for Rap2B expression associated with overall survival of patients with GBM and LGG from TCGA database. (A) Patients with GBM and high expression of Rap2B tended to have a shorter survival time compared with those in the low group, but no significant difference between two groups. (B) Patients with LGG and increased expression of Rap2B showed poorer survival outcomes compared with those with low expression. (C) No significant difference was observed between high group and low group in patients with LGG with tumor grade 2. (D) High expression of Rap2B predicted unfavorable prognosis of patients with grade 3. GBM, glioblastoma multiforme; LGG, low-grade glioma; TCGA, The Cancer Genome Atlas.
Fig 2: Rap2B activates the ERK pathway. (A) Western blot analysis showed that the protein expression of p-ERK was significantly reduced in Rap2B-silencing U87 and U251 cells. (B) Level of p-ERK was significantly increased in Rap2B-overexpressing U87 and U251 cells. Treatment with SCH772984 suppressed p-ERK expression. ***P<0.001 vs. the corresponding NC group. #P<0.05 and ##P<0.01 vs. the overexpressed Rap2B group. NC, negative control; ns, no statistical significance; p-, phosphorylated; si, short interfering; oe, overexpressed.
Fig 3: Effects of ERK pathway on Rap2B-induced MMP2 and MMP9 expression. An enzyme-linked immunosorbent assay showed MMP2 and MMP9 secretion in the supernatant of (A) U87 and (B) U251 glioma cells. Protein levels of MMP2 and MMP9 were determined by western blot analysis following treatment with Rap2B overexpression plasmids with or without SCH772984 in (C) U87 and (D) U251 cells. *P<0.05, **P<0.01 and ***P<0.001 vs. the control group. #P<0.05, ##P<0.01 and ###P<0.001 vs. the overexpressed Rap2B group. MMP, matrix metalloproteinase.
Fig 4: Effects of Rap2B on MMP2 and MMP9 expression in glioma cells. (A) After glioma cells were treated with Rap2B overexpression plasmids, an ELISA assay was employed to examine the secretion of MMP2 and MMP9. (B) Following the silencing of Rap2B, an ELISA was used to detect MMP2 and MMP9 expression in the supernatant of glioma cells. Western blot analysis and protein quantification revealed the protein levels of MMP2 and MMP9 following treatment with (C) overexpressed and (D) knocked down Rap2B. *P<0.05, **P<0.01 and ***P<0.001 vs. the corresponding NC group. NC, negative control; ns, no statistical significance; MMP, matrix metalloproteinase; si, short interfering; oe, overexpressed; ELISA, enzyme-linked immunosorbent assay.
Fig 5: Effect of Rap2B on proliferation of glioma cells. (A) CCK-8 assay showed that silenced Rap2B suppressed glioma cell proliferation. (B) CCK-8 assay showed that overexpressed Rap2B enhanced glioma cell proliferation, while SCH772984 restrained their proliferation. (C and D) An EdU assay was performed to further determine the proliferation ability of glioma cells. Magnification, ×200. *P<0.05, **P<0.01 and ***P<0.001 vs. the corresponding NC group. ##P<0.01 and ###P<0.001 vs. the overexpressed Rap2B group. NC, negative control; ns, no statistical significance; CCK-8, Cell Counting Kit 8; si, short interfering; oe, overexpressed.
Supplier Page from Abcam for Anti-RAP2B antibody