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    Signal Transduction-1

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    See attached for image.

    This figure is from Labasque et. al. 2008

    A) Describe the experiment shown here and what was learned as a result
    B) Design a negative control for this experiment and describe why it is a good control
    C) Design an experiment to confirm this result in living cells. Please include controls in your experiment design.

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    Solution Preview

    Figure 1c of the Labasque 2008 paper

    Describe the experiment that led to the result:

    From the text, for reference: (C) Solubilized protein extracts of HEK-
    293 cells transfected with empty vectors (CTL) or cotransfected with
    GFP-CaM and either wild-type 5-HT2C or 5-HT2CR376/377A receptor
    were treated with either 5-HT (1 M) or vehicle for 5 min. Solubilized
    protein extracts were immunoprecipitated with the polyclonal
    anti-GFP antibody. Immunoprecipitated proteins were analyzed by
    Western blotting using a monoclonal anti-GFP antibody and the
    polyclonal anti-5-HT2C receptor 522 antibody. Inputs represent 5%
    of the total protein amount used in immunoprecipitations. Data
    illustrated in A-C are representative of three independent experiments.

    a) In this experiment, HEK293 cells were transfected with either an empty vector ...

    Solution Summary

    In this solution, data from a real paper needed to be analyzed. The student was helped in terms of telling the major findings of a figure. In addition, the student needed help in designing an experiment to confirm the results. Two different experiments were proposed in the solution, with an external resource provided to help understand one of the concept more clearly.