Additional file 8.
Figure S4: Expression of the green fluorescent protein (GFP)-fused vertnin in cultured cells. The plasmid vector pcDNA-DEST47-VRTN, which encoded GFP-fused vertnin, was constructed from VRTN cDNA and pcDNA-DEST47 plasmid vector (Invitrogen). NIH-3T3 and HeLa cells (1 × 104 cells/chamber) were seeded on BioCoat Poly-D-Lysine 4-well Culture Slides (BD Biosciences), and then the plasmid vectors pcDNA-DEST47-VRTN and pcDNA/GW-47/CAT (Invitrogen), which encoded a GFP-fused CAT (chloramphenicol acetyltransferase) and was a control for cytoplasmic expression, were transfected into cells by using FuGENE 6 (Roche Diagnostics). Forty-eight hours after transfection, the cells were washed and the nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI) (Invitrogen). The cells were mounted with the anti-bleaching reagent DABCO (Invitrogen) and analyzed by fluorescence microscopy to examine green (GFP) and blue (DAPI) fluorescence. DAPI staining indicates the locations of nuclei, and GFP-fused vertnin has a similar expression pattern in both types of cell.
Format: PDF Size: 59KB Download file
This file can be viewed with: Adobe Acrobat Reader
Mikawa et al. BMC Genetics 2011 12:5 doi:10.1186/1471-2156-12-5