Fig. 5: Engineering the ssrA2 tag into MLKL for light-inducible necroptosis.
From: Engineering of photo-inducible binary interaction tools for biomedical applications
Scale bar, 10 μm. a Schematic illustrating inducible necroptosis mediated by ssrA2-fused MLKL-NT following its co-clustering with mCh-CRY2-sspB2-PB in the presence of photostimulation. PM plasma membrane, PB polybasic tail from STIM1, MLKL mixed lineage kinase-like domain, NT N-terminal domain (aa 1–140). b Constructs of PhoBIT2-fused MLKL and summary of light-dependent necroptosis events. c Time-lapsed confocal images of HeLa cells co-expressing the indicated constructs. The combination of A2 + B, but not A1 + B, led to the translocation of ssrA2-fused MLKL toward the PM to cause PM rupture (indicated by yellow arrows), a process visualized by Pacific Blue (blue) conjugated Annexin V staining. d Quantification of cytosolic MLKL translocation to PM (red) and Annexin V staining on PM (blue) at the indicated time points. Data are shown as mean ± s.e.m. (n = 7 or 20 cells from three biological replicates). (Source Data).
