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Plasma was purified from wild-type and germ-free C57BL/6 mouse blood using standard protocols.
NDGA was quantified in mouse blood using HPLC with electrochemical detection.
Total RNA was prepared from mouse blood using the Mouse RiboPure Blood RNA isolation kit (Ambion), and globin mRNA was removed using the Globinclear kit (Ambion).
Therefore, we sought to directly compare fibrocytes from human and mouse blood using an optimized technique, and measured functional responses in the same assay systems, thus enabling cross-species comparisons.
Here, we isolate an extremely rare population of committed MCp in naïve mouse blood using gradient centrifugation and flow cytometry with double sorting followed by culture in a myeloerythroid cytokine cocktail.
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Total RNA was obtained from 250 μL of wild-type, hemizygous, and homozygous mouse whole blood using the Mouse RiboPure Blood RNA Isolation Kit (Ambion/Applied Biosystems, Austin, TX, USA).
Platelet levels were measured by flow cytometric analysis of mouse whole blood using a FITC-conjugated GPIX antibody.
Blood glucose levels were measured on random-fed or overnight-fasted animals in mouse-tail blood using a glucometer (Elite; Bayer, Leverkusen, Germany) and serum samples were collected for insulin measurements.
Engraftment and disease progression was monitored by flow cytometric enumeration of the proportion of human versus mouse CD45+ (%hucells) cells in peripheral blood using established procedures [ 2, 10].
Depletion and recovery of Treg was monitored by flow cytometry of peripheral blood using anti mouse CD3-PerCP-Cy5.5 (BD Biosciences), CD4-e450 (eBioscience), CD25-PE (eBioscience) and FoxP3-Alexa647 (eBioscience).
Here we illustrate the unique features of QTL mapping in the DO with an example: mapping neutrophil counts in whole blood using 742 DO mice (410 female, 332 male).
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