Tofacitinib

Murine IFN-α, IFN-β, IFN-γ, IFN-λ2, and IFN-λ3, as well as human IFN-α and IFN-β were purchased from R&D Systems. For IFN-I treatment, cells were given 10 units/mL of IFN-α and IFN-β unless otherwise noted. Cells were treated with 10 ng/mL of IFN-II (IFN-γ) or IFN-III (IFN-λ2 and IFN-λ3). For JAK inhibition, cells were treated with 50 nM tofacitinib (Cayman Chemical) at the time of stimulation.

Hdac1 and Hdac2 floxed mice [21 (link)] were crossed either with villin-Cre transgenic mice [22 (link)] to ensure IEC-specific Hdac1 and Hdac2 deletion, or with villin-Cre^ER transgenic mice [23 (link)], to obtain Tamoxifen-inducible IEC-specific Hdac1 and Hdac2 deletion, in a C57BL/6J X 129SV X CD1 background. Experiments were approved by the Institutional Animal Review Committee of the Université de Sherbrooke (protocol 360-14B), according to relevant guidelines and regulations. Genotypes were determined by using selected primers to amplify genomic DNA purified with the Spin Doctor genomic DNA kit (Gerard Biotech, Oxford, OH, USA), as before [18 (link),24 (link),25 (link)]. Three- to four-month-old wild-type and IEC-specific villin-Cre Hdac1^−/−; Hdac2^−/− mice were injected intraperitoneally with 30 mg/kg of the pan-JAK inhibitor Tofacitinib (Cayman Chemical Company, Ann Arbor, MI, USA) [26 (link)], or with 10 mg/kg of the Notch inhibitor DBZ or Dimethyl Sulfoxide (DMSO), for five days (n = 3 for each group). To assess proliferation, mice were injected intraperitoneally with 10 mL/kg of bromodeoxyuridine (BrdU, Life Technologies, Grand Island, NY, USA) on day 6, for 2 h. Mice were killed on the sixth day.