Journal: Heliyon

Article Title: LncRNA CECR7 boosts hepatocellular carcinoma progression by recruiting RNA binding protein U2AF2 to enhance the stability of EXO1 mRNA

doi: 10.1016/j.heliyon.2023.e19862

Figure Lengend Snippet: CECR7 stabilize EXO1 mRNA under the mediation of U2AF2 protein. (A) The platform starBase ( http://starbase.sysu.edu.cn ) was applied to predict the potential RNA binding protein for both CECR7 and EXO1 mRNA. ( B, C ) RIP assay using antibody against U2AF2 to determine the enrichment of CECR7 and EXO1 mRNA in HCC cells (Mean ± SD; n = 3). ** P < 0.01, *** P < 0.001, Student's t -test. ( D, E ) RT-qPCR analysis and Western blot were used to detect the effect of CECR7 on U2AF2 expression. ( F, G ) U2AF2-overexpressing reversed the repression of EXO1 mRNA by CECR7 shRNA#1, and the EXO1 mRNA induction induced by CECR7-overexpressing was abrogated by U2AF2 shRNA (Mean ± SD; n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, two-way ANOVA.

Article Snippet: The protein lysates were incubated with primary antibody against U2AF2 (1:1000, NBP2-33397, Novus, USA), EXO1 (1:1000, NBP2-16391, Novus, USA) or β-actin (1:1000, ab8226, Abcam, USA).

Techniques: RNA Binding Assay, Quantitative RT-PCR, Western Blot, Expressing, shRNA

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/eLife.90993

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-U2AF2 (rabbit polyclonal) , Novus Biologicals , NBP2-04140 , WB 1:1000.

Techniques: Recombinant, SYBR Green Assay, Real-time Polymerase Chain Reaction, Labeling, Transfection, Protease Inhibitor, Immunoprecipitation, RNA Immunoprecipitation, Plasmid Preparation, Expressing, Sequencing, shRNA, Knockdown, CRISPR, Software, Gene Expression, Alternative Splicing

Journal: Journal of Immunology Research

Article Title: lncRNA ZFAS1 Promotes HMGCR mRNA Stabilization via Binding U2AF2 to Modulate Pancreatic Carcinoma Lipometabolism

doi: 10.1155/2022/4163198

Figure Lengend Snippet: Primer sequences utilized in this study.

Article Snippet: Then, 20 μ g total protein was sampled and separated on SDS-PAGE with a concentration of 10%, followed by transfer onto a PVDF membrane, 1 h of 5% defatted milk blocking, as well as overnight immersion (4°C) in corresponding primary antibodies U2AF2 (1 : 1000, Cell Signaling Technology, USA), HMGCR (1 μ g/mL, Abcam, UK), and β -actin (1 : 1000, Abcam, UK).

Techniques: Sequencing

Journal: Journal of Immunology Research

Article Title: lncRNA ZFAS1 Promotes HMGCR mRNA Stabilization via Binding U2AF2 to Modulate Pancreatic Carcinoma Lipometabolism

doi: 10.1155/2022/4163198

Figure Lengend Snippet: ZFAS1 maintains HMGCR mRNA stabilization by binding to U2AF2. (a) ZFAS1 distribution in BxPC-3 cells; (b) immunoprecipitation protein levels and RIP assay; (c) U2AF2 and HMGCR expression detected by qRT-PCT; (d) HMGCR mRNA stabilization; (e) HMGCR fold enrichment in U2AF2 relative to IgG; (f) qRT-PCR analysis of HMGCR in RIP experience; ∗ P < 0.05; ∗∗ P < 0.01; ## P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: Then, 20 μ g total protein was sampled and separated on SDS-PAGE with a concentration of 10%, followed by transfer onto a PVDF membrane, 1 h of 5% defatted milk blocking, as well as overnight immersion (4°C) in corresponding primary antibodies U2AF2 (1 : 1000, Cell Signaling Technology, USA), HMGCR (1 μ g/mL, Abcam, UK), and β -actin (1 : 1000, Abcam, UK).

Techniques: Binding Assay, Immunoprecipitation, Expressing, Quantitative RT-PCR

Journal: Journal of Immunology Research

Article Title: lncRNA ZFAS1 Promotes HMGCR mRNA Stabilization via Binding U2AF2 to Modulate Pancreatic Carcinoma Lipometabolism

doi: 10.1155/2022/4163198

Figure Lengend Snippet: Impact of U2AF2 on biological function of PC cells. (a) U2AF2 expression; (b) cell viability detection; (c) plate clonal cell formation; (d) cell invasiveness; ∗ P < 0.05; ∗∗ P < 0.01.

Article Snippet: Then, 20 μ g total protein was sampled and separated on SDS-PAGE with a concentration of 10%, followed by transfer onto a PVDF membrane, 1 h of 5% defatted milk blocking, as well as overnight immersion (4°C) in corresponding primary antibodies U2AF2 (1 : 1000, Cell Signaling Technology, USA), HMGCR (1 μ g/mL, Abcam, UK), and β -actin (1 : 1000, Abcam, UK).

Techniques: Expressing