Journal: Breast Cancer Research : BCR
Article Title: A novel role for signal transducer and activator of transcription 5b (STAT5b) in ?1-integrin-mediated human breast cancer cell migration
Figure Lengend Snippet: Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control siRNA to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b siGENOME SMARTpool oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p
Article Snippet: siRNA Transfection BT-549 and MDA-MB-231 cells were transfected with siGENOME SMARTpool siRNA targeting human STAT5b or individual custom oligonucleotides specific for STAT5a or STAT5b (siGENOME STAT5b SMARTpool duplex #3), or luciferase duplex control, all purchased from Dharmacon (Lafayette, CO).
Techniques: Expressing, Dominant Negative Mutation, Migration, Binding Assay, Multiple Displacement Amplification, Transfection, Luciferase, Construct