Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: MicroRNA-139-5p inhibits bladder cancer proliferation and self-renewal by targeting the Bmi1 oncogene.

doi: 10.1177/1010428317718414

Figure Lengend Snippet: Figure 5. MiR-139-5p inhibits c-MYC and Wnt signaling pathway via downregulation of Bmi1. (a) Expression of Bmi1 and c-MYC levels in T24 and 5637 cells was determined by western blotting. (b) Expression of Wnt signaling pathway was measured by TOP/ FOP detection in T24 and 5637 cells (*p < 0.05 compared with NC treatment (scrambled miRNA; Student’s t test)).

Article Snippet: TOP Flash (Addgene plasmid # 12456) and FOP Flash (TOP Flash mutant; Addgene plasmid # 12457) were gifted by Randall Moon.

Techniques: Expressing, Western Blot

Journal: Cancer Cell International

Article Title: P53 and Beta-Catenin Activity during Estrogen treatment of Osteoblasts

doi: 10.1186/1475-2867-5-24

Figure Lengend Snippet: Increase in beta-catenin expression during E2 treatment does not result in beta-catenin signaling through TCF/LEF response elements in ROS-PG13 cells. Cells were transfected with TopFlash (wild type promoter) and FopFlash (mutant promoter) luciferase reporters in 2% media, and E2 treatment was started three hours later. E2 treatments were staggered during the 48 h interval and all cells were harvested after 48 h after the indicated time of exposure to the hormone. Mutant FopFlash activity was unchanged during the treatment and is not shown. LiCl treatment was carried out to demonstrate the validity of the assay (Inset). Cells were exposed to LiCl or NaCl 24 h after transfection for 16 h. In both these experiments luciferase activity was measured in cell lysates using equal amounts of protein. Experiments represent average ± SEM of triplicate measurements.

Article Snippet: For reporter assays, cells were transfected with the beta-catenin responsive firefly luciferase reporter plasmids TopFlash (wild type promoter) or FopFlash (Mutant promoter) (Upstate Biotechnologies) for 48 h. Three hours after transfection, cells received 17-beta estradiol to a concentration of 10–11 M for the times indicated.

Techniques: Expressing, Transfection, Mutagenesis, Luciferase, Activity Assay

Journal: Oncotarget

Article Title: Synergistic inhibition effect of TNIK inhibitor KY-05009 and receptor tyrosine kinase inhibitor dovitinib on IL-6-induced proliferation and Wnt signaling pathway in human multiple myeloma cells

doi: 10.18632/oncotarget.17056

Figure Lengend Snippet: (A) Cell viability of RPMI8226 cells treated with IL-6 (50 ng/mL) and KY-05009 (3 μM) or dovitinib (3 μM) alone or in combination for 24 or 48 h. Data are presented as mean±SD. Experiments were performed in triplicate. (B) Relative TCF/LEF luciferase activity measured by FOPflash-normalized TOPflash luciferase activityin RPMI8226 cells treated with IL-6 (50 ng/mL) and KY-05009 (3 μM) or dovitinib (3 μM) alone or in combination for 9 h. (C) RPMI8226 cells treated with IL-6 (50 ng/mL) and KY-05009 (3 μM) or dovitinib (3 μM) alone or in combination for 1 h. The mRNA expression of indicated genes was detected by qRT-PCR analysis. (D and E) The expression of TCF4-interacting proteins and phosphorylation of TNIK detected by immunoprecipitation and Western blot of RPMI8226 cells treated with IL-6 (50 ng/mL) and KY-05009 (3 μM) or dovitinib (3 μM) alone or in combination for 9 h. * P < 0.01, * P < 0.001 versus control; # P < 0.01, ## P <0.001 versus cells treated with IL-6 alone.

Article Snippet: RPMI8226 cells were trasfected with TOPflash TCF reporter plasmid (wild-type TCF binding site) (Merck Millipore, Darmstadt, Germany), FOPflash plasmid (mutant TCF binding site) (Merck Millipore, Darmstadt, Germany), and Lipofectamine 2000 (Thermo Fisher Scientific, Boston, MA, USA) in an antibiotic-free medium.

Techniques: Luciferase, Activity Assay, Expressing, Quantitative RT-PCR, Phospho-proteomics, Immunoprecipitation, Western Blot, Control