Journal: Molecular Therapy Oncolytics

Article Title: UXT, a novel DNMT3b-binding protein, promotes breast cancer progression via negatively modulating lncRNA MEG3/p53 axis

doi: 10.1016/j.omto.2021.12.008

Figure Lengend Snippet: UXT regulated the methylation modification of MEG3 through binding to DNMT3b (A) MS-PCR assay was used to test the methylation of promoter DMR in MEG3. (B) qRT-PCR analysis of DNMT3b expression of breast cancer and adjacent nontumor tissue specimens from 13 cases of patients. (C) qRT-PCR analysis of DNMT3b expression in breast cancer cells and normal breast cells. (D) Western blot analysis of DNMT3b expression in breast cancer cells and normal breast cells. (E and F) DNMT3b mRNA and protein levels were determined using qRT-PCR and Western blot analysis. (G) The effects of DNMT3b knockdown on MEG3 expression. (H) Co-immunoprecipitation analysis on the interaction between DNMT3b and UXT. (I) The DNMT3b activity was measured in MCF7 and ZR-75-1 cells transfected with shRNA targeting UXT or control shRNA. (J) The MS-PCR was performed to determine the level of MEG3 in MCF7 and ZR-75-1 cells transfected with shRNA targeting DNMT3b or control shRNA. (K) Knockdown analysis on the interaction between DNMT3b and UXT. (L) The effects of UXT combined with DNMT3b on MEG3 levels in ZR-75-1 and MCF7 cells. ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Mouse monoclonal anti-p53 antibody (1:1,000), mouse monoclonal anti-DNMT3b antibody (1:1,500), mouse monoclonal anti-UXT antibody (1:1,000), and mouse monoclonal anti-β-actin antibody (1:2,000) were purchased from Cell Signaling Technology.

Techniques: Methylation, Modification, Binding Assay, Quantitative RT-PCR, Expressing, Western Blot, Knockdown, Immunoprecipitation, Activity Assay, Transfection, shRNA, Control

Journal: Molecular Therapy Oncolytics

Article Title: UXT, a novel DNMT3b-binding protein, promotes breast cancer progression via negatively modulating lncRNA MEG3/p53 axis

doi: 10.1016/j.omto.2021.12.008

Figure Lengend Snippet: The effects of UXT and MEG3 on nude mice xenograft After constructing the stable cells of UXT knockout combined with DNMT3b knockdown or overexpression by using matched plasmid, these cells were subcutaneously injected into the right frank of nude mice. (A and B) The image and curve of tumor volume varied with time. (C–E) qRT-PCR analysis of MEG3, p53, and DNMT3b expression levels was performed. (F) Immunohistochemistry analysis of DNMT3b and p53. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Mouse monoclonal anti-p53 antibody (1:1,000), mouse monoclonal anti-DNMT3b antibody (1:1,500), mouse monoclonal anti-UXT antibody (1:1,000), and mouse monoclonal anti-β-actin antibody (1:2,000) were purchased from Cell Signaling Technology.

Techniques: Knock-Out, Knockdown, Over Expression, Plasmid Preparation, Injection, Quantitative RT-PCR, Expressing, Immunohistochemistry