Hey,
I overexpressed a gene of interest and treated cells with a DNA-damaging agent followed by staining with Hoechst at different time points.
Many of my cells overexpressing the gene of interest got giant, horseshoe-shaped nuclei (see pictures attached). Has someone seen such a horseshoe-shaped nucleus upon treatment with a DNA-damaging agent before? What might this mean?
Cells that I transfected with the control plasmids also had giant nuclei after treatment but the number of cells with such horseshoe-shaped nuclei was lower compared to cells in which the gene of interest was overexpressed.
It is more "kidney-like" structure than "Horseshoe2 to me but the photographs do not show nothing odd in the nuclei (there is no pyknosis indicating cell death).
I think you may look for the cytoskeleton (tubulin, actin) or the Golgi for finding what is causing this modification on the nuclei structure. To me it seems something rather "physical" that makes the nuclei to accommodate.
Hei,
thank you for my answer. Yes, I have planned to stain for tubulin- I am still waiting for material I need for such experiments.
Though you have not mentioned the kind of cells that were transfceted, it is some myeloid cells (particularly of leukemic orgin), It is very likely that your gene of interest is inducing myeloid differentiation.
it`s a neuroblastoma cell line (SK-N-BE(2)).
These cells may also differentiate. However, when SKNBE(2) differentiate, they form neurite-like structures. However, my cells do not seem to differentiate.
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