Step 6
Data Analysis
Summary
Summary
The panel below compares the full length (FL) RAP6 protein and its location in transfected 293T cells. Each construct features three images: GFP filtered images corresponding to location of RAP6 protein, DAPI stains the nucleus of all fixed cells, and the third column features the merged image of both GFP and DAPI staining. It can be seen that RAP6 FL, RGD and Rab5GEF mainly localize in the cytosol of the cell. In cells transfected with the PH domain alone, Rab5 localization was observed largely in the nucleus. I want to emphasize that the figures indirectly suggest that Rab5 may be linked to the patterns of localization that we observe.
The panel below compares the full length (FL) RAP6 protein and its location in transfected 293T cells. Each construct features three images: GFP filtered images corresponding to location of RAP6 protein, DAPI stains the nucleus of all fixed cells, and the third column features the merged image of both GFP and DAPI staining. It can be seen that RAP6 FL, RGD and Rab5GEF mainly localize in the cytosol of the cell. In cells transfected with the PH domain alone, Rab5 localization was observed largely in the nucleus. I want to emphasize that the figures indirectly suggest that Rab5 may be linked to the patterns of localization that we observe.
Data
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Additional Data
Additional Data
- Compares the FL RAP6 protein expression and its localization in transfected 293T cells.
- GFP filtered images correspond to location of RAP6 protein expression.
- DAPI stains the nucleus of all fixed cells.
- MERGE contains the combined image of both GFP and DAPI staining, and is the area that is focused on in order to achieve results.
- As observed from above, RAP6 full length (FL) localizes in both the nucleus (N) and the cytosol (C) of the cell.
- Characterization of each specific domain below will provide a better idea as to where these localization signals are coming from.
- RAP6 Ras-GAP is also known as the RGD domain.
- GFP filtered images correspond to location of RAP6 protein expression.
- DAPI stains the nucleus of all fixed cells.
- MERGE contains the combined image of both GFP and DAPI staining, and is the area that is focused on in order to achieve results.
- RAP6 RGD domain mainly localizes in the cytosol of the cell.
- The GFP localization signals are seen most evident in the MERGE due to the fact that the green fluorescent protein expression is solely in the cytosol of the cell.
- RAP6 Rab5-GEF domain acts as a guanine nucleotide exchange factor thus, activating Rab5.
- GFP filtered images correspond to location of RAP6 protein expression.
- DAPI stains the nucleus of all fixed cells.
- MERGE contains the combined image of both GFP and DAPI staining, and is the area that is focused on in order to achieve results.
- It is observed that along with the Rab5 GEF domain, there is localization solely in the cytosol of the cell.
- Although not clearly observed from the MERGE images, one can depict the green fluorescent protein expression entirely around the nucleus and hence in the cytosol of the cell.
- RAP6 PH domain is generally known as the Pleckstrin Homology domain.
- GFP filtered images correspond to location of RAP6 protein expression.
- DAPI stains the nucleus of all fixed cells.
- MERGE contains the combined image of both GFP and DAPI staining, and is the area that is focused on in order to achieve results.
- RAP6 PH domain alone was observed to localize largely in the nucleus.
- Opposed to the other domains, this nuclear localization signal (NLS) is clear and thus, suggesting a possible link to the patterns that are being observed in cell proliferation (requiring further research).
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