Fig 1: The SHP-2 inhibitor PHPS1 reversed the effects of UBE2D3 knockdown. U87 and T98G cells transduced with shUBE2D3-1 or shNC and then treated with 10 µM PHPS1. (A) Expression of STAT3, and p-STAT3. (B) Cell proliferation was assessed. (C–E) Extracellular acidification rate (C), glycolytic capacity, glycolytic reserve (D) and oxygen consumption rate (E). *p < 0.05; **p < 0.01.
Fig 2: Inhibition of UBE2D3 suppressed cell proliferation and STAT3 phosphorylation as well as inducing cell apoptosis in vivo. (A, B) The volume (A) and weight (B) of subcutaneous tumors derived from mice model. U87 cells transduced with shUBE2D3-1 and NC lentivirus were subcutaneously injected in mice (n=6 per group). (C) TUNEL assay of tumors derived from mice model. Magnification, 400×. (D) The expression of UBE2D3, STAT3, p-STAT3, HK-2 and PFKL in tumors derived from mice model. **p < 0.01.
Fig 3: UBE2D3 was highly expressed in GBM and enriched in different pathways. (A) The expression of UBE2D3 in GBM and normal brain tissue. Data were extracted from TCGA-GBM dataset. (B–D) GSEA analysis of cell glycolysis (B), STAT3 pathway (C), and cell apoptosis (D) between UBE2D3 high and low group. GBM, glioblastoma multiforme; GSEA, Gene Set Enrichment Analysis.
Fig 4: UBE2D3 interacted with SHP-2 and promoted its ubiquitination. (A) The protein and mRNA levels of SHP-2 in U87 and T98G cells transduced with specific shRNAs. (B) The protein and mRNA levels of SHP-2 in U251 cells with the overexpression of UBE2D3. (C) Co-immunoprecipitation assay using lysates of U87 cells confirmed the interaction between UBE2D3 and SHP-2. (D) The expression of SHP-2 in U251 cells with overexpressed UBE2D3 in the presence of 10 µM MG132 or Vehicle (DMSO). (E) The ubiquitination and expression level of SHP-2 after the transduction of shUBE2D3.
Fig 5: Overexpressed SHP-2 reversed the effects of UBE2D3. (A) The expression of SHP-2 after transduced with SHP-2 overexpressed lentivirus in U251 cells with overexpressed UBE2D3 or not. (B, C) Cell proliferation (B) and apoptosis (C) of U251 cells with overexpressed SHP-2 and UBE2D3 or not. (D, E) Extracellular acidification rate (D), glycolytic capacity, glycolytic reserve (E) and oxygen consumption rate (F) of U251 cells with overexpressed SHP-2 and UBE2D3 or not. (G) Expression of HK-2, PFKL, STAT3, and p-STAT3 in U251 cells with overexpressed SHP-2 and UBE2D3 or not. *p < 0.05; **p < 0.01; ***p < 0.001.
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