anti-tap73 mouse mab 5b429  (Novus Biologicals)

Journal: Cancers

Article Title: Myeloid Cell Leukemia 1 Small Molecule Inhibitor S63845 Synergizes with Cisplatin in Triple-Negative Breast Cancer

doi: 10.3390/cancers15184481

Figure Lengend Snippet: Cisplatin and S63845 induce TAp73 expression and downstream targets in MDA-MB-468 cells. ( A , B ) RT-qPCR analysis of MDA-MB-468 and MDA-MB-231 cells treated over various conditions of cisplatin and S63845 for 24 h, respectively. Western blot analyses of ( C ) MDA-MB-468 and ( D ) MDA-MB-231 cells treated over various conditions of cisplatin and S63845 for 24 h. Combination treatment consisting of 100 nM cisplatin and 30 nM S63845. Differences between groups were evaluated using a matched-pair two-way ANOVA followed by Tukey’s post hoc test; p < 0.05. * p < 0.05; ** p < 0.01; *** p < 0.001. All experiments were performed in biological triplicate.

Article Snippet: All antibody dilutions for protein detection were as follows: MCL1 protein: anti-MCL1 rabbit mAb (D2W9E, Cell Signaling, Danvers, MA, USA); TAp73 protein: anti-TAp73 mouse mAb (5B429, Novus Biologicals, Centennial, CO, USA), diluted to 1:500; GAPDH protein: anti-GAPDH XP rabbit mAb (D16H11, Cell Signaling, Danvers, MA, USA); cleaved caspase 3 (CC3) protein: anti-CC3 rabbit mAb (9664S, Cell Signaling, Danvers, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancers

Article Title: Myeloid Cell Leukemia 1 Small Molecule Inhibitor S63845 Synergizes with Cisplatin in Triple-Negative Breast Cancer

doi: 10.3390/cancers15184481

Figure Lengend Snippet: MCL1 and TAp73 mediate synergistic gene expression of TAp73 and downstream targets. ( A ) Representative Western blot of MDA-MB-468 cells treated with siGFP or siMCL1. RT-qPCR analysis of ( B ) MDA-MB-468 cells transfected with siGFP or MCL1 for 24 h. RT-qPCR analysis of ( C ) MDA-MB-468 cells transfected with siGFP or MCL1 for 48 h and treated with 100 nM cisplatin for 24 h. ( D ) Representative Western blot of MDA-MB-468 cells treated with siGFP or siTAp73. RT-qPCR analysis of ( E ) MDA-MB-468 cells transfected with siGFP or TAp73 for 24 h. RT-qPCR analysis of ( F ) MDA-MB-468 cells transfected with siGFP or TAp73 for 48 h and treated with 30 nM S63845 for 24 h. Gene expression was normalized to the respective siRNA target treated with DMSO. Differences between groups were evaluated using a matched-pair two-way ANOVA followed by Tukey’s post hoc test; p < 0.05. * p < 0.05; ** p < 0.01; *** p < 0.001. All experiments were performed in biological triplicate.

Article Snippet: All antibody dilutions for protein detection were as follows: MCL1 protein: anti-MCL1 rabbit mAb (D2W9E, Cell Signaling, Danvers, MA, USA); TAp73 protein: anti-TAp73 mouse mAb (5B429, Novus Biologicals, Centennial, CO, USA), diluted to 1:500; GAPDH protein: anti-GAPDH XP rabbit mAb (D16H11, Cell Signaling, Danvers, MA, USA); cleaved caspase 3 (CC3) protein: anti-CC3 rabbit mAb (9664S, Cell Signaling, Danvers, MA, USA).

Techniques: Gene Expression, Western Blot, Quantitative RT-PCR, Transfection

Journal: Cancers

Article Title: Myeloid Cell Leukemia 1 Small Molecule Inhibitor S63845 Synergizes with Cisplatin in Triple-Negative Breast Cancer.

doi: 10.3390/cancers15184481

Figure Lengend Snippet: Figure 6. Cisplatin and S63845 induce TAp73 expression and downstream targets in MDA-MB- 468 cells. (A,B) RT-qPCR analysis of MDA-MB-468 and MDA-MB-231 cells treated over various conditions of cisplatin and S63845 for 24 h, respectively. Western blot analyses of (C) MDA-MB- 468 and (D) MDA-MB-231 cells treated over various conditions of cisplatin and S63845 for 24 h. Combination treatment consisting of 100 nM cisplatin and 30 nM S63845. Differences between groups were evaluated using a matched-pair two-way ANOVA followed by Tukey’s post hoc test; p < 0.05. * p < 0.05; ** p < 0.01; *** p < 0.001. All experiments were performed in biological triplicate.

Article Snippet: Primary Antibodies All antibody dilutions for protein detection were as follows: MCL1 protein: anti-MCL1 rabbit mAb (D2W9E, Cell Signaling, Danvers, MA, USA); TAp73 protein: anti-TAp73 mouse mAb (5B429, Novus Biologicals, Centennial, CO, USA), diluted to 1:500; GAPDH protein: anti-GAPDH XP rabbit mAb (D16H11, Cell Signaling, Danvers, MA, USA); cleaved caspase 3 (CC3) protein: anti-CC3 rabbit mAb (9664S, Cell Signaling, Danvers, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancers

Article Title: Myeloid Cell Leukemia 1 Small Molecule Inhibitor S63845 Synergizes with Cisplatin in Triple-Negative Breast Cancer.

doi: 10.3390/cancers15184481

Figure Lengend Snippet: Figure 7. MCL1 and TAp73 mediate synergistic gene expression of TAp73 and downstream targets. (A) Representative Western blot of MDA-MB-468 cells treated with siGFP or siMCL1. RT-qPCR analysis of (B) MDA-MB-468 cells transfected with siGFP or MCL1 for 24 h. RT-qPCR analysis of (C) MDA-MB-468 cells transfected with siGFP or MCL1 for 48 h and treated with 100 nM cisplatin for 24 h. (D) Representative Western blot of MDA-MB-468 cells treated with siGFP or siTAp73. RT-qPCR analysis of (E) MDA-MB-468 cells transfected with siGFP or TAp73 for 24 h. RT-qPCR analysis of (F) MDA-MB-468 cells transfected with siGFP or TAp73 for 48 h and treated with 30 nM S63845 for 24 h. Gene expression was normalized to the respective siRNA target treated with DMSO. Differences between groups were evaluated using a matched-pair two-way ANOVA followed by Tukey’s post hoc test; p < 0.05. * p < 0.05; ** p < 0.01; *** p < 0.001. All experiments were performed in biological triplicate.

Article Snippet: Primary Antibodies All antibody dilutions for protein detection were as follows: MCL1 protein: anti-MCL1 rabbit mAb (D2W9E, Cell Signaling, Danvers, MA, USA); TAp73 protein: anti-TAp73 mouse mAb (5B429, Novus Biologicals, Centennial, CO, USA), diluted to 1:500; GAPDH protein: anti-GAPDH XP rabbit mAb (D16H11, Cell Signaling, Danvers, MA, USA); cleaved caspase 3 (CC3) protein: anti-CC3 rabbit mAb (9664S, Cell Signaling, Danvers, MA, USA).

Techniques: Gene Expression, Western Blot, Quantitative RT-PCR, Transfection