Fvb n

All animal experiments were approved by and conducted under the oversight of the Massachusetts General Hospital (MGH, Boston, MA, USA) Institutional Animal Care and Use Committee (IACUC protocol number 2014N000211). Five- to seven-week-old male mice; nu/nu, FVB/n and C57BL/6 mice (Charles River Laboratories, Wilmington, MA, USA) were kept on a 12:12 light-to-dark cycle with ad libitum access to food, water, and daily health checks by Center for Comparative Medicine staff at MGH.

C57BL/6 (B6), A/J (A in the text), and C57BL/6J-Chr 10^A/J/NaJ (Css10) were purchased from The Jackson (Jax, Bar Harbor, ME) and FVB/N were purchased from Charles River Laboratories (Wilmington, MA). FVB.Ly49h transgenic (FVB.Ly49h) mice and B6.Ly49h^−/− mice were obtained as previously described [12] (link), [13] (link). A.Ly49h mice were generated by backcrossing the FVB.Ly49h mice onto an A/J WT background for a minimum of 10 generations. A genome scan using 1449 single nucleotide polymorphism showed a 100% similarity between A/J and A.Ly49h mice. Transgenic mice were identified by PCR using the Ly49h specific marker, D6Ott11 as described previously [12] (link). Recombinant congenic mice of the AcB/BcA set were derived from two successive backcrosses (N₃) to either A/J (AcB) or B6 (BcA) parental mice, as previously described [49] (link). The B6 mice deficient for H2-D^bK^b (B6.H2^−/−) were kindly provided by Dr. Hidde L. Ploegh (Cambridge, Massachusetts). The m157-Transgenic mouse was kindly provided by Dr. Sandeep K. Tripathy, (Washington University School of Medicine, St. Louis). Mice were bred and maintained in a specific pathogen-free animal facility at McGill University. All experimental protocols were developed in accordance with institutional guidelines of the Canadian Council on Animal Care.

Transgenic mice were generated as described previously by Davis et al. [9 (link)]. Briefly, the purified DNA fragment was microinjected into the pronuclei of fertilized inbred FVB/N (Charles River, Wilmington, MA) mouse eggs at the concentration of 1 ng/μl via standard techniques. Injected eggs were then surgically implanted into pseudopregnant B6CBAF1 (Charles River) foster mothers. The transgene-positive founders were mated with wild type FVB mice to confirm transmission of the transgene upon PCR analysis of tail DNAs from their offspring. To evaluate transgene expression, we performed RT-PCR targeting the oPEG11 ORF using total RNA extracted from quadriceps femoris skeletal muscles of two offspring per transmitting founder.