Assessment of Relative Expression Outline a detailed experimental design to assess the relative expression levels of Cyclin Bd1 in prostate cancer cells compared to control cells. Include the methods/assays you would employ, necessary controls, and how you would quantify the results. Discuss how you would ensure the validity and reliability of your findings.
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Question 1-2. CDK-Cyclin The temporal expression profiles for cyclins during the cell cycle are shown at right. Researchers monitored the cyclin protein expression in normal cells and cancer cells using western blots. The numbers across the top of the image represent timepoints taken every 3 hours during the cell cycle. The left set of blots show protein expression of phosphorylated Rb (pRb) and Cyclin A in normal cells. The right set of blots show protein expression of pRb and Cyclin A in cancer cells. Note: Western blot evaluation: dark bands mean more protein, light bands mean less protein! Don't worry about change in size! a. Use the information provided to identify which time points represent the G2 phase of the cell cycle in Normal Cells. Explain your reasoning. b. Cyclin A protein levels drop between zero and 9 hours in normal cells. Explain why and how cyclin A levels dropped. c. The researchers measured the kinase activity of CDK in the normal cells. What time point during the experiment would Cyclin A regulated CDK activity (phosphorylation of substrates) be high in the normal cells. Explain your reasoning. d. CDK activity was dramatically different in the cancer cells. What do you think the researchers observed when they monitored CDK activity in the cancer cells and why? e. The researchers were also measuring the amount of phosphorylated Rb protein (pRb) present in each cell type. Rb phosphorylation starts a positive feedback loop. Explain how phosphorylation of one Rb can lead to phosphorylation of many more Rb proteins.
Adi S.
1. You are working in a research lab as a summer intern. The graduate student that is supervising you wants you to determine if overexpressing the estrogen receptor protein causes an increase in p21 protein expression in HeLa cells. a. Please write the names of the experiments you would need to perform, in order, to answer this research question. A list is fine. You do not need to write each step of each experiment but please include the major reagents for the experiment. For example, if I was to include PCR, I would say I need a forward and a reverse primer for the gene and include the gene name. b. Please draw the final results of your experiment if the data show an increase in p21 protein expression when estrogen receptor is overexpressed in HeLa cells. c. Please draw the final results of your experiment if the data do NOT show an increase in p21 protein expression when estrogen receptor is overexpressed in HeLa cells.
Josee P.
In human cells, four different cyclins drive the cell cycle (see picture). Please note that the cyclins overlap (the picture shows the concentration of each cyclin protein in the cytoplasm). Write a short essay or outline of an experiment to test the following things: How can you figure out how much cyclin protein is present in the cytoplasm at different phases of the cell cycle? How can you figure out if there is overlap between the cyclins? For example, note that cyclin D overlaps with the other ones. How can you figure out which cyclin drives the cell cycle at any given cell stage? For example, cyclin E overlaps with cyclin D. What would you do to test whether cyclin E or D drives the cell into S-phase? In your answers, be as specific as you can. Use techniques that we learned in class and explain the outcomes of your proposed experiments. In other words, how would the results of your experiments test each one of the questions above? Cyclin Expression Cycle: Cyclin E Cyclin A Cyclin B Cyclin D G1 Phase S Phase G2 Phase Mitosis
Sri K.
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