Journal: Bioengineered

Article Title: Tumor suppressing role of serum-derived exosomal microRNA-15a in osteosarcoma cells through the GATA binding protein 2/murine double minute 2 axis and the p53 signaling pathway

doi: 10.1080/21655979.2021.1987092

Figure Lengend Snippet: Primer sequences for RT-qPCR

Article Snippet: The mixture was further incubated with MDM2 chromatin fragment-contained ProteinA agarose at 4°C for 2 h to conjugate protein A and antibodies.

Techniques: Sequencing

Journal: Bioengineered

Article Title: Tumor suppressing role of serum-derived exosomal microRNA-15a in osteosarcoma cells through the GATA binding protein 2/murine double minute 2 axis and the p53 signaling pathway

doi: 10.1080/21655979.2021.1987092

Figure Lengend Snippet: GATA2 mediates transcription activity of MDM2 to suppress the p53 signaling pathway. a, putative binding sites between GATA2 and the promoter region of MDM2 predicted on the UCSC website; b, binding relationship between GATA2 and MDM2 validated through the ChIP assay (* p < 0.05, two-way ANOVA); c, MDM2 expression in OS samples predicted on TCGA database (* p < 0.05, unpaired t test); d, relevance between MDM2 and prognosis of OS patients predicted on TCGA database (* p < 0.05, Kaplan-Meier analysis); e, a negative correlation between the predicted GATA2 and MDM2 expression in OS samples (* p < 0.05, Pearson’s correlation analysis); f, MDM2 expression in tumor and normal tissues from OS patients determined by RT-qPCR (n = 31, * p < 0.05, paired t test); g, relevance between MDM2 expression and the survival rate of the OS patients (* p < 0.05 according to the Kaplan-Meier analysis); h, a positive relevance between GATA2 and MDM2 expression in the tumor tissues from patients (* p < 0.05, Pearson’s correlation analysis); i, MDM2 expression in OS cell lines and in hFOB1.19 cells determined by RT-qPCR (* p < 0.05, one-way ANOVA); j, mRNA expression of MDM2 in cells transfected with MDM2-OE examined by RT-qPCR (* p < 0.05, one-way ANOVA); k, protein level of p53 in MG-63 cells after different treatment detected by western blot analysis (* p < 0.05, one-way ANOVA). Repetition = 3

Article Snippet: The mixture was further incubated with MDM2 chromatin fragment-contained ProteinA agarose at 4°C for 2 h to conjugate protein A and antibodies.

Techniques: Activity Assay, Binding Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: Bioengineered

Article Title: Tumor suppressing role of serum-derived exosomal microRNA-15a in osteosarcoma cells through the GATA binding protein 2/murine double minute 2 axis and the p53 signaling pathway

doi: 10.1080/21655979.2021.1987092

Figure Lengend Snippet: miR-15a promotes apoptosis and cell cycle arrests of OS cells. a, transfection efficiency of GATA2-OE, miR-15a mimic and MDM2-OE in MG-63 cells evaluated by RT-qPCR (* p < 0.05, one-way ANOVA); b, proliferation of cells determined by the colony formation assay (* p < 0.05, one-way ANOVA); c, apoptosis of MG-63 and U2-OS cells determined by flow cytometry (* p < 0.05, one-way ANOVA); d, protein levels of Bcl-2 and Bax in cells determined by western blot analysis (* p < 0.05, two-way ANOVA); e, cell cycle progression in cells determined by flow cytometry (* p < 0.05, two-way ANOVA); f, protein levels of CDK2, CDK4, cyclin D, cyclin E in cells determined by western blot analysis (* p < 0.05, two-way ANOVA). Repetition = 3

Article Snippet: The mixture was further incubated with MDM2 chromatin fragment-contained ProteinA agarose at 4°C for 2 h to conjugate protein A and antibodies.

Techniques: Transfection, Quantitative RT-PCR, Colony Assay, Flow Cytometry, Western Blot

Journal: Bioengineered

Article Title: Tumor suppressing role of serum-derived exosomal microRNA-15a in osteosarcoma cells through the GATA binding protein 2/murine double minute 2 axis and the p53 signaling pathway

doi: 10.1080/21655979.2021.1987092

Figure Lengend Snippet: A diagram representation of the molecular mechanism. In OS cells, miR-15a directly binds to GATA2 mRNA, whereas GATA2 promotes MDM2 transcription and suppresses the activity of the p53 signaling pathway. miR-15a suppresses while the GATA2/MDM2 axis promotes proliferation and cell cycle progression of cells. miR-15a is one of the cargos of the serum-derived exosomes and can be delivered to the cells by the exosomes

Article Snippet: The mixture was further incubated with MDM2 chromatin fragment-contained ProteinA agarose at 4°C for 2 h to conjugate protein A and antibodies.

Techniques: Activity Assay, Derivative Assay