the stretching occurred for 5 minutes before the cells were stimulated with Bradykinin. Due to the presence of EDTA, there was a different outcome observed from these experiments which is displayed in FIGURE 7 and FIGURE 8. FIGURE 7: 1% stretch with and without the presence of EDTA FIGURE 8: 5% stretch with and without the presence of EDTA As it can be clearly distinguished in the figures above, once EDTA is present, the signal intensity decreases gradually after stimulation whereas in the absence of EDTA, the Ca2+ signals drop dramatically as soon as the stimulation with Bradykinin occurs. After determining that bi-directional stretch changed the distribution of caveolae in the plasma membrane, there were experiments conducted in order to examine whether this distribution of caveolae depended on the number of neighboring cells. For these experiments, plates with very low and very high confluency of A10 cells were cultured, fixed and fluorescent labelled for caveolae. Through these experiments, it was proven that when the cells had a large amount of cell-cell contacts, the number of caveolae were much greater and the effect on calcium was more pronounced (FIGURE 9). Sample images of high and low cell-cell contact are displayed below. FIGURE 9. The distribution of caveolea in the plasma membrane depends on the confluency of the cells. Sample pictures of high density of cells (left) vs low density of cells (right) are displayed above. In addition to the images that are displayed above, Z-stack images were also captured (data is not displayed). In order to verify that the two data sets are different from each other, there was a conducted (FIGURE 10). In FIGURE 11 the box plots for the ratio of high cell-cell contact and low cell-cell contact are being displayed.
the stretching occurred for 5 minutes before the cells were stimulated with Bradykinin. Due to the presence of EDTA, there was a different outcome observed from these experiments which is displayed in FIGURE 7 and FIGURE 8. FIGURE 7: 1% stretch with and without the presence of EDTA FIGURE 8: 5% stretch with and without the presence of EDTA As it can be clearly distinguished in the figures above, once EDTA is present, the signal intensity decreases gradually after stimulation whereas in the absence of EDTA, the Ca2+ signals drop dramatically as soon as the stimulation with Bradykinin occurs. After determining that bi-directional stretch changed the distribution of caveolae in the plasma membrane, there were experiments conducted in order to examine whether this distribution of caveolae depended on the number of neighboring cells. For these experiments, plates with very low and very high confluency of A10 cells were cultured, fixed and fluorescent labelled for caveolae. Through these experiments, it was proven that when the cells had a large amount of cell-cell contacts, the number of caveolae were much greater and the effect on calcium was more pronounced (FIGURE 9). Sample images of high and low cell-cell contact are displayed below. FIGURE 9. The distribution of caveolea in the plasma membrane depends on the confluency of the cells. Sample pictures of high density of cells (left) vs low density of cells (right) are displayed above. In addition to the images that are displayed above, Z-stack images were also captured (data is not displayed). In order to verify that the two data sets are different from each other, there was a conducted (FIGURE 10). In FIGURE 11 the box plots for the ratio of high cell-cell contact and low cell-cell contact are being displayed.
Calcium_Signals_in_A10_Cells_Final_Version.docx