Next, the localization was investigated biochemically. Neurons were treated as a

Question

Next, the localization was investigated biochemically. Neurons were treated as above and then lysed under mild conditions. In these conditions, the polymerized cytoskeleton is insoluble and can be pelleted by centrifugation. Proteins in the supernatant (soluble) or pellet (insoluble) were separated by SDS-PAGE and western/immunoblotted with antibodies against: Eps8; Stg (synaptotagmin), a soluble protein; and Tubulin.

The amount of Eps8 in the soluble fraction was normalized to Stg, which was used a loading control.

Question 15

What could be interpreted from this data?

Question 16

What effect did inhibition of MEK have on Eps8?

Inhibition of MEK increased BDNF-stimulated re-distribution

Take the last two pieces of data together. What conclusions can be drawn? T/F

Question 17

When Eps8 is at the ends of the actin bundles in the filopodia, it is bound to them and insoluble.

Question 18

When Eps8 translocates away from the ends of the actin bundles into the axon, it is bound to actin and insoluble.

Explanation / Answer

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