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Donor spermatogonial lines were derived from testes of individual Germ Cell Specific (GCS -EGFP transGCS -EGFPs [31], [36].
Data from four mutant genes were analyzed: Foxa3 [32], Nebulin (neb) [33], Ercc1 [34] and Fibulin-4 (eFigure [35] (Figure 4), using RNA derived from testes, skeletal muscle, liver and aorta respectively.
While theirs was a good first approach, a logical next step would be to use biologically replicated quantitative real-time PCRs on cDNA pools derived from testes of single flies to examine the extent of misexpression.
We should add that the evidence of many of these transcripts was purely derived from testes, a tissue previously associated with unusual transcripts that appear to be non-functional or aberrant versions of functioning genes [ 40, 41].
We note the mouse and Ae. aegypti libraries were not derived from the same tissue types (the mouse libraries were derived from testes), but barring a very large difference between piRNAs in mouse testes and other mouse tissues, this should not fundamentally affect our conclusions.
We screened the expression of the previously reported neural and mesenchymal stem cell surface markers in Nes-GFP+ cells derived from testes of postnatal 7-day-old mice, including CD51, p75 (CD271), PDGFR-α, NG2, CD146, CD106, CD73, CD44, CD90, Sca-1, CD117, CD119, CD11b, and CD45 through flow cytometry.
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A study published in 2009 also claimed to have made human sperm cells in the laboratory from tissue derived from the testes, but this had to be retracted and the claims left unverified, Professor Pacey said.
Hyaluronan can be degraded by hyaluronidase and hyaluronidase derived from animal testes has been used extensively to facilitate dispersion of co-injected materials in the clinic (Bookbinder et al., 2006).
We designed primers to specifically amplify Apobec transcripts from RNA derived from the testes of the 129 mouse strain.
Primary cell cultures were derived from E13 testes, equivalent to 12 18 tail somite stage of male rat embryos, and used for promoter reporter assays.
Sea urchin ADP-ribosyl cyclase sequences were determined by PCR screening of Strongylocentrotus purpuratus cDNA libraries derived from the testes and/or ovary, the latter kindly provided by Professor Gary Wessel, Brown University, USA.
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