Tdtomato reporter mice

All mouse care and experimental procedures were approved by the institutional animal care research advisory committee of the Albert Einstein College of Medicine. Mice used in these experiments included ChAT-IRES-Cre [34 (link)] and tdTomato reporter mice (The Jackson Laboratory). 2-month-old mice on a mixed C57BL6/129SVJ background were used for all experiments. Animals were housed in groups in cages under conditions of controlled temperature (22°C) with a 12:12 h light dark cycle and fed a standard chow diet with ad libitum access to water.

C57BL/6J mice were obtained from Jackson laboratories and housed in the Boston University Animal Science Center on a 12-hour light-dark cycle with access to food and water ad libitum. Prox1-Cre-ER^T2 mice and TdTomato reporter mice were obtained from The Jackson Laboratory (strain #022075 and strain #007914, respectively). The YAP^loxP/loxP mice and Wwtr1/TAZ^loxP/loxP mice used to generate the YAP/TAZ^△LEC mice reported here were provided by Dr. Jeffrey Wrana (LTRI institute) (Jackson Laboratory Strain #030532). Our study examined both male and female animals with the sex of the mice randomized across experimental groups. Similar findings are reported for both sexes. At time of sacrifice, mice were ethically euthanized using isoflurane and inferior vena cava incision. All methods were carried out in accordance with relevant IACUC guidelines and regulations and are reported in accordance with ARRIVE guidelines. All animal experiments were performed in compliance with approved Boston University animal protocols PROTO201800710_TR01 and PROTO201800057_TR01.

Sox2-creER^T2 mice^{9 (link)}, hGFAP-cre mice^{10 (link)} and tdTomato reporter mice^{11 (link)} were obtained from the Jackson Laboratory. Nestin-GFP mice were used as previously described^{12 (link)}. Genotyping was performed by PCR analysis using genomic DNA from ear biopsies. Primers for Cre^{13 (link)} and tdTomato^{14 (link)} have been published previously.

Brain sections from 18 months old and 14 days post LPS injection C57BL/6 mice were treated with varying concentrations of copper sulfate for 1.5 h. Sections were then performed with immunofluorescence staining. To assess the effects of copper sulfate treatment on the fluorescence of RFP in a lineage tracing system, Sox10-cre/ERT2 and tdTomato reporter mice (Jackson Laboratories, Maine, ME, USA) were cross-bred. Tamoxifen was injected into the offspring, using a dosage of 1 mg per mouse per day for 3 days. The sample brains were harvested and sectioned using the vibratome method described above.

All mice used for breeding and experiments in this study were housed in a light-dark (12 h on/off; lights on at 7:00 AM) and temperature-controlled environment with food and water available ad libitum in the KAIST facilities. Npy-hrGFP mice were obtained from the Jackson laboratory (#006417). For some patch clamp experiments GIRK2 KO mice [41 (link)], used with the permission from Dr. Markus Stoffel (ETH Zurich), were crossed with Npy-hrGFP mice. Agrp-ires-Cre mice (Jackson laboratory, #012899) were crossed with tdTomato reporter mice (Jackson laboratory, #007914), GIRK1^flox/flox mice [22 (link)] or GIRK2^flox/flox mice [23 (link)] for FISH, ISH, IHC, and in vivo metabolic experiments. Mice were fed standard NCD (Teklad global 18% protein 2018S, ENVIGO).