TNF levels are increased in miR-146a–deficient T cells. (A-C) The amount of intracellular TNF in miR-146a^−/− or miR-146a^+/+ CD8⁺ T cells isolated from allo-HCT recipients on (A-B) day 7 or (C) day 3 is shown. (A) Representative flow cytometry plot gated on CD8⁺H-2Kb⁺ T cells. (B) Pooled data from 3 independent transplantation experiments. (C) Pooled data from 2 independent transplantation experiments. (D) TNF levels were measured in the serum of allo-HCT recipient mice receiving T cells from miR-146a^−/− or miR-146a^+/+ mice on day 7 after transplantation. Data were pooled from 2 independent experiments. (E) CD4⁺/CD8⁺ T cells from miR-146a^−/− or miR-146a^+/+ mice were isolated after 48 hours of stimulation with allogeneic BMDCs (BALB/c) in vitro and reached a purity of >85%. RNA was isolated from purified T cells and Tnf expression was analyzed by qRT-PCR.