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Total cellular RNA was extracted from organs using TRI Reagent (Sigma Aldrich, St. Louis, MO, USA). iNOS mRNA and the housekeeping gene GAPDH mRNA were quantified in parallel for normalization. The following 20X kits from Applied Biosystems (Foster City, CA, USA) were used for iNOS and GAPDH respectively: Mm00440502_m1 (Nos2, FAM) and Mm00484668_m1 (Gapdhs, VIC). RT-PCR was performed with Superscript III One Step Platinum Taq (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s recommendation for assay preparation and cycling.
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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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