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To determine the extent of shifting of the physiology and gene expression profiles of WAT to BAT in Fsp27−/− mice, we checked the expression levels of BAT-specific genes and several transcriptional regulators by real-time PCR analysis.
To assess whether MMPs play a role in the PNN reduction in the DCN of EE-reared mice, we checked protein expression levels and enzymatic activity of two MMPs that are expressed in the cerebellum, MMP2 and MMP9 (gelatinases) [27].
In order to confirm the expression of FoxN1 in the BM of FoxN1TguCreERT+ mice, we checked FoxN1 mRNA level with real-time RT-PCR.
In addition to the comparison between young and old mice, we checked whether IL-1α injection 24 h prior to knee joint isolation (10 ng) (R&D Systems, Wiesbaden, Germany) influenced the expression patterns.
To test the possibility that splice defects other than aberrant Clc-1 exon 7a inclusion are enhanced in Mbnl1 ΔE3/ΔE3/mice3 ΔE2 mice, we checked exon inclusion for all Clc-1 exons (Fig. S5B).
To address the mechanisms underlying metabolic disorders in Cmah-null mice, we checked the expression profiles of genes whose expressions were significantly up- or downregulated by Cmah loss in the array analysis.
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To determine the expression of AHL related genes in the cochlear tissues of Cmah-null mouse, we checked the expression profiles of these genes on the microarray analysis (Fig. 4C).
Experimentally, 106 cells were injected into the distal femur of Balb/c mice, and then we checked the K5L-Luc cell localization immediately after injection using IVIS (Fig 4A).
For each lifted mouse binding site we checked if >10% of the site was overlapped by a human site, or vice versa.
Because increased levels and activity of Brn-1 have been observed in Brn-2 knockout mouse brain (19, 25), we checked the Brn-1 protein level in R6/2 brain lysates.
As ligand-induced recombination will never be absolutely complete [ 15], we checked old Mll2FC/FC; Rosa26-CreERT2/+ mice for repopulation of various compartments with the unrecombined allele, particularly in blood.
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Justyna Jupowicz-Kozak
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