Journal: Cancer Research

Article Title: Bmi-1 Cooperates with H-Ras to Transform Human Mammary Epithelial Cells via Dysregulation of Multiple Growth-Regulatory Pathways

doi: 10.1158/0008-5472.can-07-1636

Figure Lengend Snippet: Figure 6. Analysis of p53 pathway in control MCF10A and MCF10A-derived late-passage cells. All cells except parental MCF10A cells, passage 9 cells, were used for the analysis. A, top, Western blot analysis of Bmi-1, H-Ras, and p53 in control MCF10A and MCF10A-derived (LP) cells (as indicated) was done as described in Fig. 2. Bottom, densitometric analysis of signals (of p53 and H-Ras) present in each lane was done, normalized to corresponding a-tubulin signal, and plotted to determine the expression levels of p53 and H-Ras as indicated. B, analysis of DNA damage response in MCF10A and MCF10A-derived late-passage cells. The cells were treated with camptothecin (CPT) for indicated amount of time, harvested, and analyzed by Western blot analysis for total p53, phosphorylated p53 (Ser-15 and Ser-37), p53 target genes (p21, PUMA, Bax, and PIG3), PRAK, and BCL2. h-Actin was used as a loading control.

Article Snippet: Among other antibodies, phosphorylated AKT 1/2/3 (Ser-473), AKT-1 (B-1) and AKT-2 (F-7), CDK4 (C-22), cyclin D1 (A-12), H-Ras (F-235), p21 (F-5), p53 (DO-1), p53-Ser-15, PUMA (FL-193), Bax (6A7), extracellular signal-regulated kinase (ERK; C-16), phosphorylated ERK (E-4), p38-regulated/activated protein kinase (PRAK; A-7), and QM (C-17) antibodies were obtained from Santa Cruz Biotechnology. p53-Ser-37 rabbit polyclonal antibody was obtained from Cell Signaling Technology.

Techniques: Control, Derivative Assay, Western Blot, Expressing

Journal: Cancer Research

Article Title: Bmi-1 Cooperates with H-Ras to Transform Human Mammary Epithelial Cells via Dysregulation of Multiple Growth-Regulatory Pathways

doi: 10.1158/0008-5472.can-07-1636

Figure Lengend Snippet: Figure 2. Various growth-regulatory pathways are dysregulated in cells co-overexpressing Bmi-1 and H-Ras. All cells were analyzed at passage 2 after Ras selection and/or mock infection. A, Western blot analysis of phosphorylated AKT, total AKT (AKT1 and AKT2), phosphorylated ERK, and total ERK in control MCF10A and MCF10A-derived cells (as indicated). Western blot analysis using h-actin served as a loading control. B, Western blot analysis of p53, pRb, CDK4, and cyclin D1 in asynchronously growing MCF10A and MCF10A-derived cells (as indicated). h-Actin and QM are loading controls.

Article Snippet: Among other antibodies, phosphorylated AKT 1/2/3 (Ser-473), AKT-1 (B-1) and AKT-2 (F-7), CDK4 (C-22), cyclin D1 (A-12), H-Ras (F-235), p21 (F-5), p53 (DO-1), p53-Ser-15, PUMA (FL-193), Bax (6A7), extracellular signal-regulated kinase (ERK; C-16), phosphorylated ERK (E-4), p38-regulated/activated protein kinase (PRAK; A-7), and QM (C-17) antibodies were obtained from Santa Cruz Biotechnology. p53-Ser-37 rabbit polyclonal antibody was obtained from Cell Signaling Technology.

Techniques: Selection, Infection, Western Blot, Control, Derivative Assay

Journal: Cancer Research

Article Title: Bmi-1 Cooperates with H-Ras to Transform Human Mammary Epithelial Cells via Dysregulation of Multiple Growth-Regulatory Pathways

doi: 10.1158/0008-5472.can-07-1636

Figure Lengend Snippet: Figure 6. Analysis of p53 pathway in control MCF10A and MCF10A-derived late-passage cells. All cells except parental MCF10A cells, passage 9 cells, were used for the analysis. A, top, Western blot analysis of Bmi-1, H-Ras, and p53 in control MCF10A and MCF10A-derived (LP) cells (as indicated) was done as described in Fig. 2. Bottom, densitometric analysis of signals (of p53 and H-Ras) present in each lane was done, normalized to corresponding a-tubulin signal, and plotted to determine the expression levels of p53 and H-Ras as indicated. B, analysis of DNA damage response in MCF10A and MCF10A-derived late-passage cells. The cells were treated with camptothecin (CPT) for indicated amount of time, harvested, and analyzed by Western blot analysis for total p53, phosphorylated p53 (Ser-15 and Ser-37), p53 target genes (p21, PUMA, Bax, and PIG3), PRAK, and BCL2. h-Actin was used as a loading control.

Article Snippet: Among other antibodies, phosphorylated AKT 1/2/3 (Ser-473), AKT-1 (B-1) and AKT-2 (F-7), CDK4 (C-22), cyclin D1 (A-12), H-Ras (F-235), p21 (F-5), p53 (DO-1), p53-Ser-15, PUMA (FL-193), Bax (6A7), extracellular signal-regulated kinase (ERK; C-16), phosphorylated ERK (E-4), p38-regulated/activated protein kinase (PRAK; A-7), and QM (C-17) antibodies were obtained from Santa Cruz Biotechnology. p53-Ser-37 rabbit polyclonal antibody was obtained from Cell Signaling Technology.

Techniques: Control, Derivative Assay, Western Blot, Expressing

Journal: Cancer Research

Article Title: Bmi-1 Cooperates with H-Ras to Transform Human Mammary Epithelial Cells via Dysregulation of Multiple Growth-Regulatory Pathways

doi: 10.1158/0008-5472.can-07-1636

Figure Lengend Snippet: Figure 2. Various growth-regulatory pathways are dysregulated in cells co-overexpressing Bmi-1 and H-Ras. All cells were analyzed at passage 2 after Ras selection and/or mock infection. A, Western blot analysis of phosphorylated AKT, total AKT (AKT1 and AKT2), phosphorylated ERK, and total ERK in control MCF10A and MCF10A-derived cells (as indicated). Western blot analysis using h-actin served as a loading control. B, Western blot analysis of p53, pRb, CDK4, and cyclin D1 in asynchronously growing MCF10A and MCF10A-derived cells (as indicated). h-Actin and QM are loading controls.

Article Snippet: Among other antibodies, phosphorylated AKT 1/2/3 (Ser-473), AKT-1 (B-1) and AKT-2 (F-7), CDK4 (C-22), cyclin D1 (A-12), H-Ras (F-235), p21 (F-5), p53 (DO-1), p53-Ser-15, PUMA (FL-193), Bax (6A7), extracellular signal-regulated kinase (ERK; C-16), phosphorylated ERK (E-4), p38-regulated/activated protein kinase (PRAK; A-7), and QM (C-17) antibodies were obtained from Santa Cruz Biotechnology. p53-Ser-37 rabbit polyclonal antibody was obtained from Cell Signaling Technology.

Techniques: Selection, Infection, Western Blot, Control, Derivative Assay