C57bl 6 tg cag egfp 1310sb leysopj

For all bone marrow transplant experiments, fully-acclimated (either C57 black or on a C57 black background) mice underwent lethal irradiation (950 rads divided into two sessions spaced 4 hours apart, GammaCell 40, Atomic Energy of Canada Limited, Mississauga, ON, Canada) followed by bone marrow cells (1 × 10⁷) harvested from donors. Recipients were 1) wild-type mice with 5/6 nephrectomies at 10–12 weeks salvaged with GFP-expressing marrow (C57BL/6-Tg(CAG-EGFP)1310sb/LeySopJ [stock # 006567], The Jackson Laboratory, Bar Harbor, ME), 2) CCR2-deficient mice salvaged with GFP-expressing marrow, 3) wild-type mice salvaged with CCR-deficient marrow (C57BL/6–129S4-Ccr2^tmIfc/J [004999], and C57BL/6-B6.129(Cg)-Ccr2^tm2.1Ifc/J [017586]). For the first group, partial nephrectomies were performed at 10–12 weeks followed by 2 weeks acclimation. For all groups, 4 weeks were used for engraftment, then animals were randomized into control or gadolinium contrast-treated groups (Omniscan, General Electric HealthCare, Little Chalfont, UK; 2.5 mmol/kg intraperitoneally, aiming for 20 doses over 4 weeks).

Female C57 black mice underwent 5/6 nephrectomy at 10–12 weeks. After 2 weeks acclimation, the mice were lethally-irradiated (950 rads divided into sessions spaced 4 hours apart, GammaCell 40, Atomic Energy of Canada Limited, Mississauga, ON, Canada). Bone marrow cells (1 × 10⁷) were harvested from green fluorescent protein-expressing C57 black male mice (C57BL/6-Tg(CAG-EGFP)1310sb/LeySopJ, The Jackson Laboratory, Bar Harbor, ME) and administered via the tail veins. After 4 weeks for engraftment, animals were randomized into control (n = 5) or gadolinium contrast-treated groups (Omniscan, General Electric HealthCare, Little Chalfont, UK; 2.5 mmol/kg intraperitoneally, aiming for 20 doses over 4 weeks, n = 6). All experimental procedures and protocols were in accordance with the Guide for the Care and Use of Laboratory Animals published by the NIH and approved by the Institutional Animal Care and Use Committee (IACUC). Endpoints were 1) weight loss of 10%, 2) dermatologic findings previously described (Wagner, Tan et al. 2012 (link)), or 3) completion of 4 weeks of contrast treatment. Animals were examined daily for any signs of systemic fibrosis. At the time of sacrifice, dorsal skin thickness was measured in triplicate with digital calipers (VWR, Radnor, PA).

B6.Cg-Tg-IL-2^{tm1(eGFP)Weav} (IL-2.eGFP) and B6.IL-2.BAC-inThy1.1 (2BiT) were generated using strategies previously described (23 (link)) and bred at the University of Alabama at Birmingham (UAB) animal facility. B6N-Tyr^c-Brd/BrdCrCrl (albino B6) and B6-LY5.2/Cr (congenic B6 CD45.1) were purchased from Frederick Cancer Center and intercrossed to produce albino B6.CD45.1. C57BL/6 (WT B6), Tcrb^–/– (B6.129P2-Tcrb^tm1Mom/J), OT-II (B6.Cg-Tg(TcraTcrb)425Cbn/J) mice and mice transgenic for constitutive eGFP expression (C57BL/6-Tg(CAG-EGFP)^{1310sb/LeySop}/J) were purchased from The Jackson Laboratory. SMARTA Tg (Tg(TcrLCMV)1Aox) (35 (link)) on a B6 background were a generous gift from Dr. A. Zajac (Dept. of Microbiology, UAB). All intercrosses to generate additional strains, such as SMARTA.IL-2.eGFP, SMARTA.2BiT, SMARTA.IL-2.eGFP Thy1.1⁺, OT-II.IL-2.eGFP, and 2BiT.CD45.1 were generated by crosses in UAB’s breeding facility. Animals were bred and maintained under specific pathogen-free conditions in accordance with institutional animal care and use committee regulations.