Conditional NOTCH1 Knockout in Skin Allografts

All mouse protocols were approved by the Institutional Animal Care and Use Committee of the University of Massachusetts/Amherst. Offspring between the ages of 9 and 12 weeks were used in experiments. PKCθ^null mice were obtained from Dan Littman (New York University, NY, USA). C57BL/6, p50^null, and NOTCH1 floxed mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA). NOTCH1 conditional floxed mice (N1KO) were generated by crossing NOTCH1^fl/fl (NOTCH1^tm2Rko/GridJ) to Mx1-Cre^± [B6.Cg-Tg(Mx1-cre)1Cgn/J] from the Jackson Laboratory (Bar Harbor, ME, USA). To conditionally delete NOTCH1 from splenocytes of skin allograft recipients, poly(I)–poly(C) (Amersham Biosciences, Piscataway, NJ, USA) was dissolved in phosphate buffered saline (PBS) and given in vivo as previously described (23 (link)).