Journal: Cell Death & Disease

Article Title: The RBPJ/DAPK3/UBE3A signaling axis induces PBRM1 degradation to modulate the sensitivity of renal cell carcinoma to CDK4/6 inhibitors

doi: 10.1038/s41419-022-04760-6

Figure Lengend Snippet: a Bioinformatic analysis of the potential binding proteins of the promoter of DAPK3 via analysis the ChIP-seq datasets. b , c 786-O and ACHN cells were transfected with indicated plasmids for 24 h. Cells were harvested for western blotting analysis ( b ) and RT-qPCR analysis ( c ). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SEM with three replicates ( n = 3). Ns not significant; ** P < 0.01; *** P < 0.001. d , e 786-O and ACHN cells were infected with indicated shRNAs for 72 h. Cells were harvested for western blotting analysis ( d ) and RT-qPCR analysis ( e ). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SEM with three replicates ( n = 3). ** P < 0.01. f , g 786-O cells were treated with 0, 1 µM, 5 µM, and 10 µM the RBPJ inhibitors (the DAPK3 inhibitor) for 24 h. The WCL of cells were subjected to western blotting analysis ( f ) and RT-qPCR analysis ( g ). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SEM with three replicates ( n = 3). Ns not significant; *** P < 0.001. h the ChIP-seq of RBPJ on the promoter region of DAPK3 . i 786-O cells were infected with indicated shRNAs for 72 h. Cells were harvested for ChIP-qPCR analysis by using the IgG or RBPJ antibodies. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SEM with three replicates ( n = 3). ** P < 0.01; *** P < 0.001. j 786-O cells were transfected with indicated plasmids for 24 h. Cells were harvested for ChIP-qPCR analysis by using the IgG or RBPJ antibodies. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SEM with three replicates ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001. k The correlation between RBPJ and DAPK3 were analyzed by the GEPIA web tool ( http://gepia.cancer-pku.cn/ ) in different types of cancer. l 786-O cells were infected with indicated shRNAs for 72 h. Cells were harvested for western blotting analysis. m 786-O cells were infected with indicated shRNAs for 48 h. Then, cells were treated with or without the 5 µM RBPJ inhibitor for another 24 h. Cells were harvested for western blotting analysis.

Article Snippet: The antibodies used as follows: UBE3A (10344-1-AP, Proteintech; 1:1000 dilution), PBRM1 (12563-1-AP, Proteintech; 1:500 dilution), DAPK3 (2928, Cell signaling technology, 1:1000 dilution), RBPJ (14613-1-AP, Proteintech; 1:1000 dilution); P21 (10355-1-AP, Proteintech; 1:1000 dilution); GAPDH (10494-1-AP, Proteintech; 1:10000 dilution).

Techniques: Binding Assay, ChIP-sequencing, Transfection, Western Blot, Quantitative RT-PCR, Infection, ChIP-qPCR

Journal: Cell Death & Disease

Article Title: The RBPJ/DAPK3/UBE3A signaling axis induces PBRM1 degradation to modulate the sensitivity of renal cell carcinoma to CDK4/6 inhibitors

doi: 10.1038/s41419-022-04760-6

Figure Lengend Snippet: DAPK3 competed with PKA to bind with UBE3A and enhance the PBRM1 degradation in renal cancer cells. PBPJ transcriptionally regulated DAPK3 expression and then promoted UBE3A-mediated degradation of PBRM1. Then, PBRM1 increased the p21 expression and sensitized renal cancer cells to CDK4/6 inhibitors. In combination with RBPJ inhibitors, CDK4/6 inhibitors synergistically enhanced renal cancer cells.

Article Snippet: The antibodies used as follows: UBE3A (10344-1-AP, Proteintech; 1:1000 dilution), PBRM1 (12563-1-AP, Proteintech; 1:500 dilution), DAPK3 (2928, Cell signaling technology, 1:1000 dilution), RBPJ (14613-1-AP, Proteintech; 1:1000 dilution); P21 (10355-1-AP, Proteintech; 1:1000 dilution); GAPDH (10494-1-AP, Proteintech; 1:10000 dilution).

Techniques: Expressing

Journal: iScience

Article Title: Molecular docking-aided identification of small molecule inhibitors targeting β-catenin-TCF4 interaction

doi: 10.1016/j.isci.2021.102544

Figure Lengend Snippet:

Article Snippet: Notch CSL reporter , BPS Bioscience , Cat# 60652.

Techniques: Recombinant, Luciferase, Cell Viability Assay, CCK-8 Assay, RNA Sequencing Assay, Negative Control, Mutagenesis, Plasmid Preparation, Software, High Content Screening