Results
14 – 3 -3 sigma gene produces 4N cell having an irregular shape 14 – 3 – 3 sigma gene was overexpressed to determine its function in tumor cells. The amount of living cells for G was 81% while for the S sample, 73% was measured. This unexpected minor difference may be due the stress caused while handling the sample e.g. the incubator was opened several times causing fluctuation in temperature, essential gases (e.g. oxygen) and the cells were taken out of the incubator for transfection, seeding and addition of nocodazole.
As 14 – 3 – 3 sigma (S) was found to be a tumor suppressor, arresting cells in the G2 or M phase of the cell cycle3, it`s hypothesized, overexpression of S leading to cells containing more 4N cells than in G. …show more content…
As tubulins, are features associated with shape and mitotic spindle it`s hypothesized, 14 – 3 – 3 sigma`s arrest interferes with the spindle formation and thereby causing changes in shape. To test this, IFA assay was performed.
While inspecting the transfected cells in G (figure 7) and S, it was found that indeed the spindle seems to be irregular (figure. 8), likely altering the shape.
Nocodazole may impair mitotic spindle formation
To investigate the effect of nocodazole on the mitotic spindle an IFA staining was performed. Nocodazole interferes with the spindle formation during the mitosis, therefore, it is speculated, treated cells having a defective spindle. Figure 9 and 10 show that nocodazole is indeed able to impair spindle formation as cells treated with nocodazole showed an irregular spindle shape.
Presence of nocodazole in 14 – 3 – 3 sigma enhances impaired spindle formation
As 14 – 3 – 3 sigma and nocodazole both, cause spindle deformation, it`s hypothesized, the amount of impaired spindles being larger in treated cells containing the 14 – 3 – 3 sigma transfection. The experiment, demonstrated more spindle deformations in S (figure 11) compared to G (figure 10), confirming the
14 – 3 -3 sigma gene produces 4N cell having an irregular shape 14 – 3 – 3 sigma gene was overexpressed to determine its function in tumor cells. The amount of living cells for G was 81% while for the S sample, 73% was measured. This unexpected minor difference may be due the stress caused while handling the sample e.g. the incubator was opened several times causing fluctuation in temperature, essential gases (e.g. oxygen) and the cells were taken out of the incubator for transfection, seeding and addition of nocodazole.
As 14 – 3 – 3 sigma (S) was found to be a tumor suppressor, arresting cells in the G2 or M phase of the cell cycle3, it`s hypothesized, overexpression of S leading to cells containing more 4N cells than in G. …show more content…
As tubulins, are features associated with shape and mitotic spindle it`s hypothesized, 14 – 3 – 3 sigma`s arrest interferes with the spindle formation and thereby causing changes in shape. To test this, IFA assay was performed.
While inspecting the transfected cells in G (figure 7) and S, it was found that indeed the spindle seems to be irregular (figure. 8), likely altering the shape.
Nocodazole may impair mitotic spindle formation
To investigate the effect of nocodazole on the mitotic spindle an IFA staining was performed. Nocodazole interferes with the spindle formation during the mitosis, therefore, it is speculated, treated cells having a defective spindle. Figure 9 and 10 show that nocodazole is indeed able to impair spindle formation as cells treated with nocodazole showed an irregular spindle shape.
Presence of nocodazole in 14 – 3 – 3 sigma enhances impaired spindle formation
As 14 – 3 – 3 sigma and nocodazole both, cause spindle deformation, it`s hypothesized, the amount of impaired spindles being larger in treated cells containing the 14 – 3 – 3 sigma transfection. The experiment, demonstrated more spindle deformations in S (figure 11) compared to G (figure 10), confirming the