Journal: Scientific Reports
Article Title: Simultaneous knockdown of uPA and MMP9 can reduce breast cancer progression by increasing cell-cell adhesion and modulating EMT genes
Figure Lengend Snippet: Cells were allowed to migrate in the transwell chamber under chemotactic condition and the migrated cells were stained with DAPI and counted under fluorescence microscope. ( A ) Bargraph showing comparative migratory ability of the MDA-MB-231, MCF-7, and T47D cells ( B ) MDA-MB-231 cells were transfected with uPA and MMP9 siRNA either singly or in combination and allowed to migrate through the membrane of the transwell unit for 18 hrs under chemotactic condition. Migrated cells were stained with DAPI. The photomicrographs were taken under fluorescence microscope using UV filter ( C ) The migrated cells were counted under fluorescence microscope ( D ) Cell after transfection with respective siRNAs were allowed to invade through the matrigel coated membrane of the transwell unit for 22 hrs and stained with DAPI. Photomicrographs were taken under fluorescence microscope using UV filter ( E ) The number of the invaded cells was then counted. Control indicates the cells were not transfected with either of the uPA and MMP9 siRNA. (Data are representative of three independently performed experiments and are expressed as mean ± SD).
Article Snippet: siRNA transfection Predesigned siRNA against uPA and MMP9 were procured from Ambion by life technologies, The sequences of siRNA against uPA was: 5′-GCUUAACUCCAACACGCAAtt-3′ (S) and 5′-UUGCGUGUUGGAGUUAAGCct-3′ (AS) and while siRNA against MMP9 was 5′-GACCU GGGCAGAUUCCAAAtt-3′(S) and 5′-UUUGGAAUCUGCCCAGGUCtg-3′ (AS).
Techniques: Staining, Fluorescence, Microscopy, Multiple Displacement Amplification, Transfection