Rosa26idtr

Mice expressing diphtheria toxin receptors (DTR) in Kupffer cells were produced by crossing Clec4f-cre-tdTomato mice to Rosa26-Lox-Stop-Lox-DTR mice (Rosa26-iDTR, The Jackson Laboratory, Stock No.007900) (Buch et al., 2005 (link)) and depleted of Kupffer cells by a single intraperitoneal administration of 200ng Diphtheria toxin from Corynebacterium diphtheriae (Sigma Cat#D0564) 8 weeks before sorting liver macrophages. Clec4f-cre-tdTomato-positive DTR-negative mice were used as control. All mice were fed CDAHFD diet (Research Diets, A06071302) (Matsumoto et al., 2013 (link)) for 4 weeks prior to sorting. Upon harvesting, plasma was saved for multiplex assessment of an inflammatory cytokine panel. Whole liver pieces were processed for histology and hematoxylin and eosin stained sections were scored by a board certified pathologist for steatohepatitis using the NASH CRN scoring system (Kleiner et al., 2005 (link)).

C57BL/6 mice 8–12 weeks of age were obtained from Charles River (USA).
As previously described (1) the Fate⁺ model was obtained by crossing the original Foxp3^RFP IL-10^eGFP IL-17A^Kata mice with IL-17a^CRE+/+ R26YFP^+/+ mice (kindly provided by Prof B. Stockinger). The iFate mice were generated in our Lab (Gagliani, Amezcua Vesely et al. 2015 (link)). The iFate mice were used in heterozygosis for the IL-17a^{IRES-eGFP-CRE-ERT2} allele (Gagliani, Amezcua Vesely et al. 2015 (link)).
ROSA26iDTR were acquired from the Jackson laboratories (007900) and crossed with IL17^{CRE +/+} FoxP3 RFP^+/+ R26STOP^flox/flox YFP^+/+ to obtain the Fate-DTR mice.
Heterozygous IL7GFP/+ mice (Miller C. N. et al, Int Immunol 2013 (link)) were provided Joao Pereira. Animal procedures were approved by the Institutional Animal Care and Use Committee of Yale University

C57BL/6, Tac1^−/− vGlut2^flox, ROSA26-tdTomato and ROSA26-iDTR mice were obtained from Jackson laboratories (Bar Harbor, ME). Mcpt5-Cre+ Rosa26 DTA (referred to MC^DTA) mice were generously provided by Axel Roers (University of Technology Dresden, Dresden). Mrgprb2^−/− mice were kindly provided by Benjamin McNeil (Northwestern University, Chicago). Mrgpra3^cre mice were generously provided by Xinzhong Dong (Johns Hopkins University, Baltimore). Human Langerin-DTA (referred to LC^DTA), Human Langerin-DTR (referred to LC^DTR), Itgb6^−/−/Itgb8^ΔKC and Mrgprd^cre ROSA26-tdTomato (referred to Mrgprd^TdT) mice have been previously described (Bobr et al., 2010 (link); Kaplan et al., 2005 (link); Mohammed et al., 2016 (link); Rau et al., 2009 (link)). Mrgprd^cre and Mrgpra^cre mice were crossed with ROSA26iDTR mice to obtain Mrgprd^DTR and Mrgpra3^DTR mice, respectively. We crossed LC^DTA mice with MC^DTA mice to obtain LC^DTAxMC^DTA mice. We generated Mrgprd^TdT mice with LC^DTA mice to obtain Mrgprd^TdTxLC^DTA mice. Mrgprd^cre mice were bred with vGlut2^flox mice to obtain Mrgprd^ΔvGlut2/+ mice. Experiments were performed on independent cohorts of male and female mice between the ages of 6-10 weeks. All mice were maintained under specific-pathogen-free conditions and all animal experiments were approved by University of Pittsburgh Institutional Animal Care and Use Committee.