Fig. 2

FOLE-induced necrotic cell death is dependent on RIP1. a Representative immunoblots of RIP1 and RIP3. IEC-6 cells were treated with FOLE at 0.25–1%, and the indicated proteins were analyzed at 8 h after FOLE treatment. b Cell viability. IEC-6 cells were pre-treated with Nec-1 (20 μM) for 40 min, followed by exposure to 0.25–1% FOLE as described in material and methods. The viability was analyzed 8 h after treatment. Data are presented as mean ± SD. *, p < 0.05, **, p < 0.01 (n = 6–8 wells/group). c Representative immunoblots of RIP1 and RIP3. IEC-6 cells were pre-treated with Nec-1 (20 μM) for 40 min, followed by exposure to 0.5% FOLE. The indicated proteins were analyzed at 2–8 h after treatment. d Representative images of RIP1 and RIP3 staining in IEC-6 cells. Cells were treated as described in (c). Scale bar = 50 μm. Three experiments were performed that showed similar results