FIGURE 1
What are we dealing with? Tip-links connect two stereocilia = mechanical sensitivity
- it is risky to include the model at the beginning because it can lead to circular logic but it can also help us as readers to understand the paper better (image k)
- why use guinea pig because they have lower frequency hearing + longer stereocilia, b/c w/ higher frequency hearing you have a lot shorter stereocilia
- it is easier to identify locations with longer stereocilia
- if you break the tip link, the CDH23 will move up, this is one way to prove that the cadherin is part of the TL
- they can predict the position of each of the antibodies from the base of the lower stereocilia
- b/c you know the sequence and repetition, you can predict the distance b/w the EC domains
- the experimental results are somewhat far from the expected distances for images f-h, is this good enough? yes
- you are req’d to provide a SD, you can do this by providing a bar graph with SD error bars (for images f-h) but it is better to
- there are 40 measurements falling between 1-20 nm and if you average all of them together get 37 for instance
FIGURE 2
- His tag used to purify the protein,
- there is a homodimer and multiple strands to anchor into the membrane
- there is better agreement
- why is there such a small difference compared to the experimental results they show in Figure 1
- it is in a much controlled environment, you already expressed the protein, and purified it
- image 1 is in the live cell and can have different tension b/c of position in stereocilia which gives them different lengths
- what about orientation, it looks like 100% have parallel arrangement of filaments, homodimers
- the evidence looks very convincing but of the 195, only 131 were on one end
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