Poly 1 poly c

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)).

The conditional Jak2^V617F mice were previously described (13 (link)). Conditional transgenic IDH1^R132H mice (expressing a mutation in the IDH1 gene) were provided by Kwok-Kin Wong (Dana-Farber Cancer Institute), and conditional Idh2^R140Q mice were provided by Craig B. Thompson (Memorial Sloan Kettering Cancer Center). All mice were maintained on a C57BL/6J background, and the lines were backcrossed for 6 generations before use in experiments. Samples for experiments were obtained from male and female mice aged 6 months to 1.5 years. Upon induction, mice received 5 i.p. injections of polyI:polyC (Amersham) of 200 μl of a 1-mg/ml solution. Peripheral blood was collected via cheek bleeding using heparinized microhematocrit capillary tubes (Thermo Fisher Scientific). Peripheral blood counts were obtained using a HemaVet according to the manufacturer’s instructions. Excision 2 weeks after induction was confirmed using PCR.