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In vitro samples were incubated under osteogenic conditions and in vivo samples were implanted subcutaneously into severely compromised immunodeficient mice, for 4 weeks.
In this study, in vitro samples were investigated prospectively.
Total RNA of in vivo and in vitro samples were purified by using two different methods.
In vitro samples were acquired by using a FACSCalibur flow cytometer and Cell Quest software (BD).
The CD4+, CD8+ T cells, granulocytes and in vitro samples were finally corrected by subtracting the number of reads in each 30 bp window by the number of reads in the control falling in the same window.
In that study, we demonstrated that at the level of backbone nodes (i.e., the differential network backbone values), the in vivo and in vitro samples were significantly correlated [ 23].
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Here, a novel method for the measurement of glycolysis reactants from in vitro samples is presented.
The genes that showed the greatest increase in the in vivo samples when compared to the in vitro samples are listed in figure S1.
The assay for hTERT mRNA expression of the in vitro samples was conducted in the same manner as for the human blood samples described above.
Phosphopeptides from the in vitro sample were enriched using TiO2 beads (GL Sciences, MZ-Analysentechnik) similar to the method described previously [ 31].
For LC ESI MS/MS analysis, 14 μl of the in vivo sample, 10 μl of the phosphopeptide fraction and 5 μl of the non-phosphopeptide fraction (in vitro sample) were injected.
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