Question

2) As a continuation of the experiments in Question 1 you want to study the timing of the APC's activity more distinctly. To do this you treat your cells with MG132- a proteasome inhibitor. When MG132 is added to cells proteins can still be ubiquitinated, but they are not degraded, because the proteasome is inhibited (4pts) In cells that are treated with MG132 you can monitor APC activity on Cyclin B using just the cyclin B antibody a. and a western blot, and monitoring the size of cyclin B. Describe how and why this works. b. (8 pts) You take a group of cells and synchronize them so that they are all transitioning through the cell cycle simultaneously. At 4 different transition times you remove some cells, treat them with MG132 and allow them to (attempt to) undergo the transition. You then run a western blot using your cyclin B antibody. Below is the start of the Western Blot that you would put in your paper. Complete this blot, and write a figure legend to accompany the figure. Be sure to add all relevant labels to the figure and write the figure legend you would expect to find in the paper. The cells are allowed to transition into the final stage listed, so G1Swould look like cells in the S phase, unless those cells got stuck somewhere along the way. Remember MG132 inhibits ALL of the proteasome activity. Figure 1 MG132 (3pts) Using your knowledge of the cell cycle, the cyclins and the APC explain the results you drew above. C.

Hi! I am having trouble answering these questions, more specifically I am stuck on part B! Could you please help me out and let me know how to answer this?

Thanks!

Answer 1

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