foxo3a knockdown  (Cell Signaling Technology Inc)

Journal: Breast cancer research and treatment

Article Title: Simvastatin prevents triple-negative breast cancer metastasis in pre-clinical models through regulation of FOXO3a

doi: 10.1007/s10549-015-3645-3

Figure Lengend Snippet: (A) Immunoblot with anti-pFOXO3a and anti-FOXO3a antibodies in SUM 159, MDA-MB-231, MCF 7, and SUM 149 cells treated with 2.5μM simvastatin or vehicle control. Cells were treated with EGF 10 ng/mL for 15 minutes following 24 hours of simvastatin treatment or DMSO treatment. Beta-actin was used as a loading control. (B) A model of how simvastatin regulates FOXO3a.

Article Snippet: FOXO3a Knockdown and Overexpression SignalSilence siRNA targeting FOXO3a and nontargeting siRNA were purchased from Cell Signaling.

Techniques: Western Blot, Control

Journal: Breast cancer research and treatment

Article Title: Simvastatin prevents triple-negative breast cancer metastasis in pre-clinical models through regulation of FOXO3a

doi: 10.1007/s10549-015-3645-3

Figure Lengend Snippet: (A) Western blot assay to identify the overexpression efficiency of the plasmids for FOXO3a in SUM 159 cells. (B) Mammosphere formation presented as the average number of spheres per 40,000 cells plated relative to the control plasmid ± SD; representative data are shown. (C) Representative images of the Transwell assays (magnification, × 40). Quantification of the numbers of migrating cells relative to the control plasmid is presented as mean ± SD. (D)Immunoblot with anti-FOXO3a antibodies in SUM 149 cells transfected with either siRNA targeting FOXO3a or scrambled control siRNA for 48 hours. Beta-actin was used as a loading control. (E) SUM 149 cells transfected with either siFOXO3a or scrambled siControl exposed to simvastatin (2.5 μM) treatment (24 h) and seeded in self-renewal promoting suspension culture conditions. Mammosphere formation presented as the average number of spheres per 40,000 cells plated ± SEM; representative data are shown. (F) SUM 149 cells transfected with either siFOXO3a or scrambled siControl migrated through an 8 μM pore in the presence of 2.5 μM simvastatin or vehicle (DMSO). The number of cells that passed through the transwell membrane was counted. Significant differences are shown as follows: *, P < 0.05 for unpaired 2-tailed Student’s t-test (n = 3).

Article Snippet: FOXO3a Knockdown and Overexpression SignalSilence siRNA targeting FOXO3a and nontargeting siRNA were purchased from Cell Signaling.

Techniques: Western Blot, Over Expression, Control, Plasmid Preparation, Transfection, Suspension, Membrane

Journal: Breast cancer research and treatment

Article Title: Simvastatin prevents triple-negative breast cancer metastasis in pre-clinical models through regulation of FOXO3a

doi: 10.1007/s10549-015-3645-3

Figure Lengend Snippet: Kaplan-Meier curves showing low FOXO3A mRNA levels (red curves) were associated with shorter metastasis-free survival in all breast cancers (top, left), in TN breast cancers (top, right), and in HR+/ERBB2− breast cancers (bottom, left). No correlation is found in ERBB2+ breast cancers (bottom, right).

Article Snippet: FOXO3a Knockdown and Overexpression SignalSilence siRNA targeting FOXO3a and nontargeting siRNA were purchased from Cell Signaling.

Techniques: