All mice used in the current study were housed in the same rooms in Emory’s animal facility during the course of the experiment, with one room designated for animal breeding and maintenance and an adjacent room solely for SE experiments. The lights were on a 12-h on/off cycle, and mice were fed and watered ab libitum. In Ccr2rfp/rfp KO mice, the Ccr2 open reading frame was disrupted with a cDNA encoding red fluorescent protein (RFP)20 (link). The Ccr2rfp/rfp mice originally obtained were maintained on the C57BL/6 (Jackson Laboratories) inbred genetic background, and we subsequently backcrossed the mice to C57BL/6 (Charles River) for over 11 generations. To generate heterozygous Ccr2+/rfp mice, in which Ccr2-expressing monocytes are marked by RFP expression, male Ccr2rfp/rfp mice were bred to female C57BL/6 mice from Charles River.
After backcrossing for over 11 generations we submitted genetic material obtained from a Ccr2rfp/rfp mouse to Charles River Laboratories for strain background characterization. A total of 128 single nucleotide polymorphisms (SNPs) spread across the genome was analyzed to generate an allelic profile for comparison between the Jackson Laboratories and Charles River C57BL/6 substrains. Our Ccr2rfp/rfp mice were a 99.6% match to C57BL/6 inbred mice from Charles River Laboratories, making the mouse strain essentially congenic.
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