p53 responsive luciferase reporter pg13 luc  (Addgene inc)

Journal: International Journal of Molecular Sciences

Article Title: p53, miR-34a and EMP1—Newly Identified Targets of TFF3 Signaling in Y79 Retinoblastoma Cells

doi: 10.3390/ijms20174129

Figure Lengend Snippet: TFF3 overexpression enhances p53 transcriptional activity and protein expression and increases miR-34a expression with downstream reduction of EMP1 in Y79 RB cells. ( A ) Quantitative Real-time PCR confirmation of TFF3 lentiviral overexpression (Trefoil factor family peptide 3 (TFF3)) in Y79 cells compared to control cells (ctr). ( B ) Luciferase assays were performed with Y79 cells transiently transfected with TFF3 or empty vector control (ctr) in addition to wild-type PG13-Luc (wt PG13) or mutant MG15-Luc (mut MG15). Forced TFF3 expression leads to an increased luciferase signal upon p53 promotor activation in Y79 cells. ( C ) Western blot analysis showing increased p53 and TFF3 protein levels after TFF3 overexpression (TFF3). The indicated intensity ratios of p53 and TFF3 protein levels relative to β-actin levels were calculated using ImageJ software. ( D , E ) Quantitative real-time PCR analysis of miR-34a and EMP1 expression levels in Y79 cells compared to control cells after lentiviral TFF3 overexpression (ctr). Values are means of at least 3 independent experiments ± SEM. * p -value < 0.05, ** p -value < 0.01; statistical differences compared to the control group calculated by Student’s t -test or one-way ANOVA and Newman-Keuls post test.

Article Snippet: TFF3 dependent p53 activity was measured with the p53 responsive reporter PG13- Luc (with p53 binding site) and the non-responsive reporter MG15- Luc (with a mutant p53 binding site; Addgene [ ]) after co-transfection of pCS2+_UBQ_TFF3 or an empty control vector (pCS2+_UBQ).

Techniques: Over Expression, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Control, Luciferase, Transfection, Plasmid Preparation, Mutagenesis, Activation Assay, Western Blot, Software

Journal: International journal of cancer

Article Title: Reduction of the tumorigenic potential of human retinoblastoma cell lines by TFF1 overexpression involves p53/caspase signaling and miR-18a regulation.

doi: 10.1002/ijc.30768

Figure Lengend Snippet: Figure 3. TFF1 overexpression enhances p53 transcriptional activity and protein expression and reduces miR

Article Snippet: TFF1 dependent p53 activity was measured with the p53 responsive reporter pG13Luc (with p53 binding site) and the non-responsive reporter pG15-Luc (with a mutant p53 binding site; Addgene,19) after co-transfection of pCS2+_UBQ_TFF1 or an empty control vector (pCS2+_UBQ).

Techniques: Over Expression, Activity Assay, Expressing