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NE102 Lab Write-Up #2

So wherever the dark circles were in image a we saw a

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DAPI that the cells were dyed with. So wherever the dark circles were in image (a), we saw a blue circle appear in image (b). Image (c) is an image of the cells that were treated with NGF for one hour. There is a clear difference because the green fluorescent color is more dominant than in the cells that were not treated with NGF. This proves that NGF has increased the presence of Egr1. Furthermore, image (d) shows the nuclei of the cells in image (c). When we switched back and forth between the two images, we could see that the bright green circles in image (c) lined up with the nuclei in image (d). This proves our theory about the localization of Egr1: because it is a transcription factor, it should be found in the nuclei of cells. We did the same analysis with the NFT L cells that were treated and untreated with NGF. Figure 2 displays our results for the NFT L cells. Image (a) shows the NFT L cells without treatment of NGF. As shown in the image, there is almost nothing that appears and that is because NFT L is a protein that is found in neurites. However, PC12 cells that are not treated with NGF will not differentiate into a neuron and therefore will not have any neurites. But we are sure that the cells still exist because of the DAPI image to the right of it, image (b). We clearly see blue circles, which signify the nuclei of cells
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8 so we know that there are in fact cells present. Images (c) and (d) illustrate the NFT L cells that were treated with NGF. As compared to image (a), image (c) is greener, which shows that there is NFT L present and that its levels increased with the addition of NGF. Again, we see darker circles in the middle of the green fluorescent parts of the cell and comparing those with image (d) we concluded that they were the nuclei of the cells. The green fluorescent color in image (a) comes from the neurites that grew off of the nuclei with treatment of NGF. This proves that NFT L is found in the neurites because NFT L was the protein that the antibody was binding too. The two figures prove our hypotheses to be correct. Treatment of cells with NGF will increase their levels of Egr1 and NFT L. Also, the ICC helped us to prove the localization of these proteins: Egr1 does in fact appear in the nucleus and that NFT L is found in the neurites of differentiated PC12 cells.
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