i did mechanical compression on cancer cells and then looked at actin staining pattern. I lack experience and did not see any changes in actin pattern.
it could take a keen eye to detect changes in the distribution of actin filaments and i am hoping for advise!
first top image is compressed actin pattern
below image is uncompressed actin pattern
Can you provide some extra information? What is the cell type? Were they grown in 3D culture or monolayer? How did you label the actin, phalloidin or antibody? Are these living cells or fixed cells when the image was taken?
The reason I am asking is that some of your methodology could be masking the changes that you seek.
Hi thanks for answering.
These are mda-mb-231 cells grown as a confluent monolayer on transwell insert before mechanical weight compression is applied on it. In between the weight and the cells are a layer of thin agarose(with media dmem composition) thus making it a 3D envuronment.
After mechanical compression i did immunofluoresece on the transwell membrane. Fixing with paraformaldehyde before doing phalloidin staining (alexa 546 nm)
I guess you dont see any changes too?
I guess I thought that your images were live cell images. I am not sure what I would expect now that I know they are fixed images.
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