.(4 pts) EphB receptor is a tyrosine kinase found in neurons and many other cell
ID: 213011 • Letter: #
Question
.(4 pts) EphB receptor is a tyrosine kinase found in neurons and many other cell types. It is known to associate with GRIP-1. We wanted to see if inhibition of GRIP-1 would affect EphB protein levels so we did a Western blot. We loaded equal amounts of protein into each well for control neurons and neurons exposed to RNAi for GRIP-1. Based on the Western blot data, does inhibition of GRIP-1 affect EphB protein levels and if so how is it affecting them? (Lane 1-control, Lane 2-RNAi for GRIP-1) D. (4 pts) We wanted to examine the effect GRIP-1 inhibition would have on EphB receptor in a different way so we did immunofluorescence on the neurons above using a primary antibody against EphB and a secondary antibody that would fluoresce green. Based on these results, what can you say about where EphB should be located at in the neuron normally and what happens in the absence of GRIP-1. . Control RNAExplanation / Answer
Answer 1. As we can see in the western blot, in both the lanes protein band is equal suggesting that the expression of EphB is similar in both the cases. Therefore it means that there is no effect of RNAi mediated inhibition of GRIP-1 on the expression levels of EphB protein.
Answer 2. Based on the results seen in the immunofluorescence results we can say that EphB should be located on the outer surface of the neuron in normal cases and when GRIP-1 is inhibited then the protein somehow changes its location to intermembrane space of the neuron as can be seen in the picture. So we can say that GRIP-1 inhibition changes the localization of EphB receptor in neurons.
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