Hi All,
I am trying to understand my mitochondria mass data. The cells are overexpressing a protein that make them smaller in size. TEM shows not real obvious change in mitochondrial size or number between control and overexpression cells. However, when we perform the standard QPCR assay looking at total genomic and mitochondrial DNA, it indicates a significant increase in mitochondrial DNA in the cells that overexpress our protein. This does not appear to be the case just looking at all the TEM images. Anyone have any ideas or thoughts on this observation?
Hi!
If you did not see any differences in the the total amount of mitochondrium by cell or mitochondrial size, shape, etc...
Did you try differents target genes/regions for mitochondrial DNA? May be a change in the copy number of mtDNA, or in the recombination rate in mtDNA that could be modifying the rate mtDNA/nDNA
I hope this help you!
try staining the cells in fixed volume with mitotracker, wash then, measure signal intensities and divide by the number of cells. staining vary from cell type to cell type but yours seem to be isogenic so it should not be a problem
Thanks for the two recommendations!!! I will try what Natanael Mansilla suggests first since by TEM we see not differences and I am not sure how staining the mito with mitotracker would change what we see with TEM. Keep the suggestions and thoughts coming!!!!
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