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Signal Transduction-1

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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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.

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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.

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