Hello and welcome back!
The results are in! And they’re consistent—something I hadn’t
realized was so important to experiments until now. After putting the U266 (the
cell line that reacted to the drug, even though we didn’t expect it to) through
a drug titration (an experiment to see at what concentration the cells are
affected) it showed marked signs of inhibited growth when a drug concentration
of 1 uM was added. This is good news, since it not only verifies that we are
following the drug company’s protocol but also that I am not horrible at lab
experiments.
Unfortunately for me, I must’ve done too well—because this
week I had to extend the drug titration to all four cell lines. That means 16
different drug concentrations, 160 wells, and 800,000 cells. My poor thumbs.
We’ve also begun to look at the protein and RNA synthesis of
the treated cell lines. The ASO’s are meant to decrease production of IRF4 by
inhibiting its RNA production. So, we should expect to see a marked decrease in
RNA and protein production in cell lines treated with the active drugs. It will
also be interesting to see the results of the cell lines that did not react to
the drug. It may give us a clearer insight as to why the drug has no effect. We
will be using a western blot to determine protein production (something I’ve
heard about in Biology class but have never tried). It’s bound to be
interesting!
Until then, thank you for reading!
Angela
Sounds like you have your work cut out for you. Do you know what kind of antibodies you're going to be using for the blot?
ReplyDeleteAha......practical experience! It's so good that you are showing your lab expertise. It's exciting to translate theoretical information into practice. Thumbs up.
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