Figure 5
Translocation of the N-terminal four-α-helix bundle of MLKL to plasma membrane is necessary and sufficient for triggering necroptosis, and targeting to plasma membrane is determined by the certain sequence within the α-helix bundle. (A) Schematic representation of full-length and truncated ND-HBD*-3×HA. The corresponding positions of the different deletions in α-helices are shown in the right panels. (B) Viabilities of MLKL KO L929 cells expressing ND-HBD* or truncated ND-HBD* (11-190, 21-190, 41-190, 1-130) as described in A after stimulation with or without 4-OHT (1 μM) for 2 h. Data represented the mean ± SD of triplicates, and were representative of three independent experiments. (C) Expression levels of ND-HBD* and its truncated forms in cells described in B were determined by immunoblotting with the anti-HA antibody. (D) MLKL KO L929 cells reconstituted with full-length or truncated ND-HBD*-3×HA were treated with or without 4-OHT (1 μM) for 2 h and then immunostained with anti-HA antibody and counterstained with DAPI and Mitotracker Red. Each scale bar represents 10 μm.
