Question

I need help with a and b please.

4) Based on the image below (from Dean et al 2001), answer the following questions (7). Serine/threonine protein phosphatase 5 (PP5) Control Nuclear > Cytoplasmic Localization of GR-GFP (%) ISIS 18321 Dermethasone Control Dexamethasone Dexamethasone Figure 2: 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 PPS), 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. a. What affect does Dexamethasone have on GR? Justify by highlighting data that supports this observation (2). b. What is the effect of treating the cells with ISIS 15534, Justify by highlighting data that supports this observation (2)?

Answer 1

GR - GFP is a protein. The given condition is studying about the localisation of this protein in the cell. The localisation is also studied by using different types of chemical which may increase or decrease the localisation of this protein in the cell.

A. There is a positive effect of dexamethasone on the subcellular localisation of this protein. This is because under the controlled condition, when dexamethasone is not there in the medium, only around 25% of the protein molecules were localised but when dexamethasone was added then around 90% of the protein molecules were localised in the cell.

B. There is a positive effect of ISIS on the subcellular localisation of this protein. This is because in the presence of ISIS around 100% of the protein molecules got localised into the cell.

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