Hello all!
Well, the main question is in the title...
I have been observing that the migration of my cells (perivascular cells) is enhanced when I treat them with a molecule. At the same time, adhesion is reduced (which isn't too surprising, as I may reach the optimum strength of attachment that allows this phenomenon). However, I observe a drastic decrease in FAK and especially paxillin phosphorylation and total protein expression... consistent with the reduction of cell adhesion, but not with migration. I performed scratch assay as well as transwell migration assay to assess migration.
Any theory that could light me up?
Thank you so much.
Carole
Hi Carole,
Probably, in normal conditions basal (high) adhesion level in your cell system favour the stickiness of the cells limiting their migration. When you induce a decrease in adhesion points, the cells less become less "sticky" (but still maintaining a threshold level of adhesion enough to support migration) and therefore migration is now promoted.
Check the link below
https://www.nature.com/articles/377539a0
Hi Carole,
Do you observe these differences by Western blot? Perhaps trying to assess the localization of these proteins (and phospho) would help you figure out what is going on. You might be losing a subset of adhesions that are required for stronger attachment (i.e. fibrillar adhesions) but not more dynamics focal adhesions.
http://jcs.biologists.org/content/120/1/137
Hi Colin and Sebastian and thank you.
Yes Colin, I observe these differences by WB. For p-FAK Y397, I also observed a loss by immunostaining.
I agree that a reduced adhesion might enhance cell migration, but I wouldn't have expected such a huge decrease in FAK and especially paxillin. My concern is how to explain this biological effect from a mechanistic point of view, as the role of FAK and paxillin in migration is preponderant.
Also, the cells are fine and alive! They just display a drastic change in their morphology, but otherwise, they absolutely don't die :)
Could also be a different mode of migration then. What does the actin staining look like? Is it organized in stress fibers? Strong cortical staining?
Also, check out Margaret Frame's papers. She's done lots of work on the functional impact of loss of FAK.
" Could also be a different mode of migration then " +> my thought, too!! I will check Frame's papers, thank you so much :)
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