Effects of ISIS 15534 and dexamethasone on the subcellular distribution of GR-GFP.A. Representative subcellular distribution of GR-GFP. A549 cells were grown and treated with ISIS 15521 (a mismatched control antisense oligonucleotide for ISIS 1535), ISIS 15534 (antisense targeting PP5), dexamethasone (500 nM), ISIS 15534 and 500 nM dexamethasone, or vehicle alone (control) as described in Material and Methods. Eighteen hours later, GR-GFP expressing plasmids were microinjected into the nuclei of cells, incubated for 6 hours in serum-containing medium, and then serum-starved for an additional 18 hours. Between 150 and 200 cells were injected for each condition. The cells were either left untreated (control, ISIS 15521, ISIS 15534) or incubated with 500 nM dexamethasone (dexamethasone, ISIS + dexamethasone) for 30 minutes, prior to fixing and observation by fluorescence microscopy. Representative cells from 4 independent experiments are shown. B. Percentage of cells displaying nuclear GR-GFP. Cells from 4 independent experiments were scored for their intracellular distribution of GR-GFP and the mean percentage of cells having greater fluorescence in their nuclei than cytoplasm are shown (± SD). *p ≤ 0.0001 by ANOVA vs untreated or mismatch-treated cells.
Dean et al. BMC Cell Biology 2001 2:6 doi:10.1186/1471-2121-2-6