All animal experiments described in this article conform to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the University of Delaware Institutional Animal Care and Use Committee. All mice were maintained and bred under specific pathogen free conditions at the University of Delaware animal facility. Wnt reporter mice (STOCK Tg(TCF/Lef1-HIST1H2BB/EGFP)61Hadj/J ), harboring a canonical Wnt reporter gene consisting of six copies of the T cell factor (TCF)/lymphoid enhancer-binding factor (Lef) binding site upstream of a hsp68 minimal promoter driving the expression of a histone 2b/green fluorescent protein (GFP) fusion protein (cWnt-nlGFP mice) (Ferrer-Vaquer et al., 2010 (link)), were obtained from the Jackson Laboratory (Bar Harbor, Maine) and crossed to wild-type (C57BL/6J, Stock #000664) mice (Jackson Laboratory). Embryos were obtained from timed matings, with noon on the day that the copulatory plug was found defined as embryonic day (E) 0.5. For genotyping, DNA was isolated from tail snips using the PureGene Tissue and Mouse Tail kit (Gentra Systems, Minneapolis, MN) followed by PCR analysis using the primers and protocol recommended by Jackson Laboratory (https://www2.jax.org/protocolsdb/f?p=116:5:0::NO:5:P5_MASTER_PROTOCOL_ID,P5_JRS_CODE:29499,013752 ).
C57bl 6j stock 000664
Manufactured by Jackson ImmunoResearch
C57BL/6J (Stock 000664) is a mouse strain commonly used in biomedical research. It is a fully inbred strain that has been maintained by brother-sister mating for over 20 generations. This strain is widely used as a control strain in various research studies.
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6 protocols using c57bl 6j stock 000664
1
Wnt Reporter Mice for Developmental Studies
2
Adipose-specific DRP1 Knockout Mice Cold Tolerance
3
Mouse Strain Maintenance and Housing
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C57BL/6J (Stock #000664), FoxP3 RFP (C57BL/6-FoxP3 tm1Flv/J, Stock #008374 mice, all on the C57BL/6 background were purchased from the Jackson Laboratory (Bar Harbor, ME). Mice were housed in a 12-h light/dark cycle specific pathogen free facility and fed standard chow (Purina 5010) ad libitum. Enrichment and privacy were provided in mating cages by “breeding huts” (Bio-Serv S3352-400). Mouse studies and all animal housing at Case Western Reserve University were approved by and performed according to the guidelines established by the Institutional Animal Care and Use Committee of CWRU.
4
Calpastatin Transgenic Mouse Model
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Mice overexpressing calpastatin (CAST-Tg) driven by a cytomegalovirus promoter on a C57BL/6 background were generated originally in the laboratory of Dr. Laurent Baud49 (link). CAST-Tg mice were backcrossed at least 9 generations into a C57BL/6 background. C57BL/6 J (Stock# 000664) mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Age-matched controls (8–10 weeks old) were used for the present study. Mice were maintained in a barrier facility. All study procedures were approved by the University of Kentucky Institutional Animal Care and Use Committee (Protocol # 2011–0907). This study followed the recommendations of The Guide for the Care and Use of Laboratory Animals (National Institutes of Health).
5
Acclimating C57Bl/6J Male Mice
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10–11 weeks old C57Bl/6J (Stock# 000664) male mice were obtained from Jackson Laboratories (Bar Harbor, ME). The animals were acclimatized to their new environment for a week and had continuous access to food and water. During the acclimatization period and throughout the experiment, the animals were housed on a reversed light cycle and were handled daily by the laboratory staff. The reverse light cycle was used because mice are nocturnal animals, so the behavioral measures were made during their active periods. Daily handling was done to habituate animals to laboratory staff and handling procedures, thereby reducing any stress associated with handling the animals during the experimental procedures.
6
Conditional knockout mice for metabolic studies
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All of the animal studies were reviewed and approved by the Animal Welfare Committee (AWC) of University of Texas Health Science Center at Houston (Animal protocol number: AWC-21-0019). Wild-type (WT) C57BL/6J (Stock 000664) and Adiponectin-Cre (Stock 010803) mice were purchased from Jackson Laboratories. The Ces1d-floxed (Ces1dflx/flx) mouse was generated by Dr. Grant A Mitchell from the Ste-Justine Hospital in Montreal, Canada, and crossed onto C57BL/6J background (18 (link)). 8-wk-old male FKO and their littermate floxed control (from here on referred to as WT) mice were used for all the experiments. Mice were housed in the animal facility (22°C ± 1°C) on a 12-h light/dark cycle with ad libitum access to water and regular chow diet, unless indicated otherwise. For the HFD challenge, the age matched male WT and FKO mice were fed with the HFD (containing 60% calories from fat; Bio-Serv) for 14 or 25 wk. The total body mass, fat mass and lean mass were measured by EchoMRI-100T (Echo Medical Systems).
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