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5 protocols using b6 c h2 ab1bm12 khegj bm12

1

Genetic Manipulation of Murine Immune Cells

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Seven- to 8-week-old WT C57BL/6J (C57BL/6 or WT, The Jackson Laboratory, stock no. 00064), B6(c)-H2-Ab1bm12/KhEgJ (BM12, The Jackson Laboratory, stock no. 001162), B6.129S2-Cd28tm1Mak/J (CD28KO, The Jackson Laboratory, stock no. 002666), BALB/cByJ (BALB/c, The Jackson Laboratory, stock no. 001026), B6.129S7-Rag1tm1Mom/J (RAGKO, The Jackson Laboratory, stock no. 002216), B6N.Cg-Tg(ACTFLPe)9205Dym/CjDswJ (FLPe B6N, The Jackson Laboratory, stock no. 019100), and B6.Cg-Tg(Cd4-Cre)1Cwi/BfluJ (CD4-Cre, The Jackson Laboratory, stock no. 022071) mice were used. PI3Kγ−/− C57BL/6 mice (PI3KγKO, backcrossed 11 generations) were received from Bao Lu (Boston Children’s Hospital/Harvard Medical School, Boston, Massachusetts, USA) and maintained in our animal facility (126 (link)).
Pik3cgtm1a(EUCOMM)Wtsi (Pik3cg-tm1a) mice were purchased from Wellcome Trust Sanger Institute (Hinxton, United Kingdom). Male and female mice were housed in sterilized and ventilated cages in a specific pathogen–free animal facility under a standard 12-hour light/12-hour dark cycle. Mice were fed irradiated food and water ad libitum. All experiments were performed with age- and sex-matched, 8- to 12-week-old mice in accordance with the relevant guidelines and regulations approved by the IACUC of Brigham and Women’s Hospital.
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2

Murine Model for HUVEC Cell Research

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HUVEC cells (CRL-1730™) were purchased from ATCC®. C57BL/6J (JAX#000664) and B6(C)-H2-Ab1bm12/KhEgJ (BM12, JAX#001162) were obtained from the Jackson Laboratory. Male or female mice were used at 7–8 weeks of age and were housed in sterilized and ventilated cages in a specific pathogen-free animal facility under a standard 12 hours light/12 hours dark cycle. Mice were fed irradiated food and water ad libitum. Each individual experiment was performed using three to four mice per group. All animal experiments and methods were performed in accordance with the relevant guidelines and regulations approved by the Institutional Animal Care and Use Committee of Brigham and Women’s Hospital, Boston, MA.
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3

Chronic GVHD Induction and Treatment

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TC mice have been described previously (22 (link)). C57BL/6J (B6), B6(C)-H2-Ab1bm12/KhEgJ (Bm12), and B6.129P2-Tcrbtm1Mom/J (TCRβ KO) mice were originally bought from the Jackson Laboratory. Only female mice were used in this study. Chronic graft- vs. host-disease (cGVHD) was induced in 8–10 weeks-old B6 mice by intravenous injection of 5 × 107 splenocytes from Bm12 mice. For in vivo treatment, mice were randomly divided into two groups and gavaged with CG-5 (100 mg/kg per mouse per day) or vehicle alone (0.1% Tween 80 and 15% dimethyl sulfoxide in water). CG-5 was obtained from Ohio State University. All experiments were conducted according to protocols approved by the University of Florida Institutional Animal Care and Use Committee.
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Transgenic Mouse Models for PD-1 Signaling

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WT C57BL/6J (B6; stock no: 000664), B6(C)-H2-Ab1bm12/KhEgJ (bm12; stock no: 001162), BALB/cJ (stock no: 000651), and C3H/HeJ (stock no: 000659) mice were purchased from The Jackson Laboratory. Foxp3-GFP mice were a gift from the Alexander Y. Rudensky laboratory (Memorial Sloan Kettering Cancer Center) (50 (link)). Descriptions of PD-1–KO (51 (link)), PD-L1–KO (29 (link)), and T cell–specific PD-1 Tg mice (12 (link)) have been published previously. Briefly, in the PD-1 Tg mice, PD-1 is overexpressed under the hCD2 promotor (12 (link)). PD-1 Tg mice were crossed with Foxp3-GFP animals in our animal facility. All mice were at 8–12 weeks of age and were harbored and used following the Harvard Medical School and National Institutes of Health guidelines.
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5

Transgenic Mouse Models in Immunological Research

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C57BL/6J (B6) (RRID: IMSR_JAX:000664) WT female mice were from Charles River. B6(C)-H2-Ab1bm12/KhEgJ (bm12) (RRID: IMSR_JAX:001162) female mice were from the Jackson Laboratory. B6.129P2-Cd38tm1Lnd/J (Cd38−/−) (RRID: IMSR_JAX:003727) female mice were backcrossed for 12 generations to the C57BL/6J (B6) background and were bred and maintained under specific pathogen-free conditions at the IPBLN-CSIC Animal Facility in Granada, Spain. The experimental procedures in animals at IPBLN-CSIC, Spain, were approved by the Institutional Animal Care and Use Committee. The procedures follow the ARRIVE guidelines (20 (link)) in accordance with the U.K. Animals (Scientific Procedures Act, 1986) and associated guidelines (EU Directive 2010/63/EU for animal experiments) and with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications No. 8023, revised 1978).
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