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The mice were split into two groups.
To determine if the GR QTL were sex-specific, we compared the number of QTL found separately in males and females to the number found when the same set of 370 mice were split into random halves, irrespective of their genders.
Here, WT and T- Atg7 −/− mice were split into three groups.
Briefly, all mice were split into either control (water) or ethanol (EtOH) group (n = 15/group).
For SYK inhibitor treatment studies, 2 weeks after transplantation, mice were split into two similar groups (seven to eight mice per group) according to the GFP content in the peripheral blood and body weight.
Mouse ears from 3-4 weeks old female AKR mice were split into two and incubated in 0.5% Dispase II (Boehringer Mannheim, Germany) in phosphate buffered solution (PBS) (Gibco BRL) at 37oC for 1 hour.
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Ankle joints dissected from the left hind leg of each mouse were split open longitudinally through the midline between the tibia and the talus, prefixed overnight with 2.5% glutaraldehyde in phosphate buffer, pH 7.4, and rinsed with the buffer.
Macrophages from each mouse were split into three groups and left as non-activated controls, classically activated by supplementing medium with 100 ng/ml INF-γ (Sigma) and 100 ng/ml LPS (Sigma) or alternatively activated by supplementing the medium with 50 μg/ml anti-INF-γ (Bio X Cell) and 20 ng/ml IL-4 (Peprotech) for 24 hours.
Indeed recently profiles of microbially-derived metabolites were reported to show considerable variability over time in healthy adult mice [31] and in another study populations of mice that were split into separate housing facilities showed changes in community fingerprints over time that correlated with their environment [32].
All mice of the same strain were co-housed in a filter top cage with the exception of the IRF9 KO mice which were split into two cages (total of 5 cages for the four strains).
In order to limit the number of blood draws per mouse, groups were split in two and blood was taken only once a day for each mouse (either at 2 hours or 12 hours post DPPIV inhibitor administration).
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