Fig. 3: LINC00998 associates with CBX3 and prevents CBX3 ubiquitination degradation.
From: LncRNA LINC00998 inhibits the malignant glioma phenotype via the CBX3-mediated c-Met/Akt/mTOR axis
a Mass spectrometry analysis of CBX3 binding to LINC00998. b, c RNA immunoprecipitation assay in A172 and U251 cells confirmed the interaction of CBX3 and LINC00998. d Coexpression analysis showed a strong correlation between CBX3 and LINC00998 in TCGA database (R = 0.54). e Fluorescence in situ hybridization combined with immunofluorescence assays demonstrated the colocalization of LINC00998 and CBX3 in the cell nucleus. f mRNA levels of CBX3 were not changed after LINC00998 overexpression in A172 and U251 cells or knockdown in U373 cells, as detected by qRT-PCR. g Protein expression of CBX3 increased or decreased after LINC00998 overexpression in A172 and U251 cells or knockdown in U373 cells, respectively. h CBX3 downregulation by LINC00998 overexpression was reversed by treatment with the proteinase inhibitor MG-132 (5 µM, 24 h). i, j Western blotting results showed that LINC00998 overexpression stabilized CBX3 protein, while LINC00998 knockdown shortened the half-life of CBX3 after CHX treatment at 20 µg/ml for 0, 2, 4, 8, and 10 h. k, l IP assays showed that LINC00998 overexpression decreased CBX3 ubiquitination levels, while LINC00998 knockdown increased ubiquitination levels. (*P < 0.05, **P < 0.01, ***P < 0.001).
