The results of deletions of Pix, Git, Fat, and expanded on the D. Melanogaster’s eyes and wings showed how disruption of these proteins affected the Hippo (Hpo) pathway and ultimately growth regulation. Following these results, what effects on cell growth would happen if Hpo was mutated so that it could not function was tested. To do this, wild type fly eyes were compared to flies that exhibited the HpoMGH1 mutation. The results showed that the flies in which Hpo exhibited loss of function showed a slight increase in number of cells compared to wild type (Figure 3A and 3B). To see how this mutation would affect cell growth and density in fly eyes if coupled with either a loss of function of Pix or Git or a loss of function of both. The combination
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The over expression of LacZ produced not a normal sized wing and eye acting as a control (Figure 4I and 4O). The over expression of both Pix and Git did not show any noticeable increase or decrease in the size of the fly eye or wing (Figure 4J and 4P). There was a slight decrease in wing size but the fly eye seemed to be normal when the overexpression of LacZ was paired with Hpo (Figure 4K and 4Q). The most noticeable affect occurred when there was an over expression of Hpo, Pix, and Git. The results were a significant decrease in the size of the fly eye and wing showing that there was minimal cell growth (Figure 4L and 4R). This significant decrease in size of wing and eye was even smaller than doubling up on the concentration of over expressed Hpo, which was still smaller than any of the other mutations (Figure 4M and 4S). Figure 4N demonstrates the decrease in wing area depending on the mutation, with the overexpression of Hpo, Pix, and Git being the smallest. This shows that the regulation pathway can be reversed to decrease tissue size if over …show more content…
To do so, two cells with DAPI (blue) and added Venus fluorescent proteins (Green) were tagged. These proteins are only detectable when there is a dimerization of Hpo. The first cell, which just included the Hpo kinase- dead proteins (HpoVc-VN), exhibited no green fluorescence meaning that Hpo did not dimerize (Figure 4A). The second cell, which included Pix and Git as well as of HpoVc-VN, showed a large amount of green fluorescents (Figure 4B). This means that Pix and Git are necessary to form the Hpo dimer. Figure 4C shows the relative levels of the fluorescent intensity meaning the higher the intensity the more Hpo dimers there are in the cells. Hpo with Pix and Git as well as Hpo, Sav with Pix and Git exhibited the much more fluorescents than any of the other cells since both Pix and Git were present in this cell as well as Hpo (Figure 4C). The phosphorylation of Hpo at its activation loop (T195) when Pix and Git were present alone and together where the tested. Western blots in which Hpo, Pix, and Git were tagged revealed that the largest quantity of phosphorylated Hpo was present when both Pix and Git were present (Figure 4D). The phosphorylation of Hpo also occurs directly by Pix and Git and not through any other means since the most amount of phosphorylated Hpo
The over expression of LacZ produced not a normal sized wing and eye acting as a control (Figure 4I and 4O). The over expression of both Pix and Git did not show any noticeable increase or decrease in the size of the fly eye or wing (Figure 4J and 4P). There was a slight decrease in wing size but the fly eye seemed to be normal when the overexpression of LacZ was paired with Hpo (Figure 4K and 4Q). The most noticeable affect occurred when there was an over expression of Hpo, Pix, and Git. The results were a significant decrease in the size of the fly eye and wing showing that there was minimal cell growth (Figure 4L and 4R). This significant decrease in size of wing and eye was even smaller than doubling up on the concentration of over expressed Hpo, which was still smaller than any of the other mutations (Figure 4M and 4S). Figure 4N demonstrates the decrease in wing area depending on the mutation, with the overexpression of Hpo, Pix, and Git being the smallest. This shows that the regulation pathway can be reversed to decrease tissue size if over …show more content…
To do so, two cells with DAPI (blue) and added Venus fluorescent proteins (Green) were tagged. These proteins are only detectable when there is a dimerization of Hpo. The first cell, which just included the Hpo kinase- dead proteins (HpoVc-VN), exhibited no green fluorescence meaning that Hpo did not dimerize (Figure 4A). The second cell, which included Pix and Git as well as of HpoVc-VN, showed a large amount of green fluorescents (Figure 4B). This means that Pix and Git are necessary to form the Hpo dimer. Figure 4C shows the relative levels of the fluorescent intensity meaning the higher the intensity the more Hpo dimers there are in the cells. Hpo with Pix and Git as well as Hpo, Sav with Pix and Git exhibited the much more fluorescents than any of the other cells since both Pix and Git were present in this cell as well as Hpo (Figure 4C). The phosphorylation of Hpo at its activation loop (T195) when Pix and Git were present alone and together where the tested. Western blots in which Hpo, Pix, and Git were tagged revealed that the largest quantity of phosphorylated Hpo was present when both Pix and Git were present (Figure 4D). The phosphorylation of Hpo also occurs directly by Pix and Git and not through any other means since the most amount of phosphorylated Hpo