I see lots of vacuoles kind of shape in the cytoplasm of my transfected cells, where I transfected it with a mutant protein. However, I don't see this pattern in Wild type transfection.
What could be the underlying mechanisms? which experiments you recommend to be done
Yes, a protein mutation can induce cytoplasmic vacuolization and lead to cell death. This can happen when the mutated protein is not able to perform its normal functions, leading to a build-up of molecules in the cytoplasm, which can lead to vacuolization and eventually cell death.
Hi Nawara,
You can also check if this mutation induces autophagy, so maybe you are observing autophagosomes, you can check it measuring LC3 through Western blot or Flow Cytometry approaches.
There are a lot of things that lead to an imbalance in cytoplasmic ion content. We tested many kinds of inhibitors, like ouabain, amiloride, valinomycin, etc. The most potent was monensin. It was not published but it is well-known that ionophores can do this. I think our article describes related data "Enhancement of the transformed shape phenotype by microtubule inhibitors and reversal by an inhibitor combination. C A Heckman, H K Plummer, R Mukherjee: April 1, 2000 https://doi.org/10.3892/ijo.16.4.709". It may be available on ResearchGate.
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