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To test the applicability of our method for archival tissue, we extracted 11-month old FFPE mouse liver using the pressure-assisted protocol developed for our model systems.
The mMCSF cDNA fragment was cloned from mouse liver using primers for the cloning into the NdeI-XhoI sites of the expression vector pACYCDuet-1 (Novagen), and the resulting plasmid was designated pSS110.
This methodology was demonstrated by performing ChIP on hepatocytes isolated directly from mouse liver using well-characterized antisera against the liver-enriched transcription factor Hnf4α [7], [15, [15] and the trimethylated lysine 4 of histone H3 [3], [5], [12] [12].
Genome-wide assay of transcription in the mouse liver using Nascent-Seq.
Overexpression of lipasin in mouse liver using adenovirus dramatically increased serum TAG levels [ 17].
Interestingly, we detected Pax8 staining in the developing mouse liver using the polyclonal antibody (Fig. 2c, f, i).
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Immunoblot analysis with anti-Dkk-1 antibodies showed a band corresponding to the molecular size of Dkk-1 (35 kDa), which was prominent in the mouse liver, used as a control tissue.
Total RNA was prepared from cell cultures and mouse livers using RNABee™ (Biozol).
We reconstructed the miRNA-mediated regulatory network in mouse livers using high-throughput expression profiles.
Total RNA was extracted from mouse livers using TRIzol reagent (Invitrogen, Breda, The Netherlands), and purified and DNAse treated using the SV Total RNA Isolation System (Promega, Leiden, The Netherlands).
Total RNA was isolated from frozen mouse livers using Trizol Reagent (Invitrogen Corporation, Carlsbad, CA) and further purified using the RNeasy Mini Kit (Qiagen Inc., Valencia, CA).
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mouse blood using
mouse placentae using
mouse tissue using
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mouse liver following
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