Fig. 7

SMART monitors poly(I:C)-induced necroptosis. a HaCaT cells were stimulated with the combination of the indicated agents for 24 h. Cell viability was determined by WST assay. Results are mean ± s.d. of triplicates. Statistical significance was determined using the one-way ANOVA test. ***P < 0.001. Results are representative of two independent experiments. b–d HaCaT cells stably expressing hSMART were stimulated with Poly(I:C) + BV6 + zVAD (PBZ) or Poly(I:C) + BV6 + GSK’872 (PBG). The FRET/CFP ratio was analyzed as in Fig. 1b. Maximum changes of the FRET/CFP ratio (b). Each dot indicates individual cell (n = 10 cells). Statistical significance was determined using the unpaired two-tailed Student-t test. ***P < 0.001. Representative images of the ratio (left) and kinetics of ΔFRET/CFP (right) of a single cell (n = 10 cells) (c, d). Red arrowheads indicate SYTOX-positive cells. Time 0 indicates the time of cells that became SYTOX-positive. Scale bar, 20 μm. Color scales indicate pseudocolor images of the FRET/CFP ratio. Error bars indicate s.e.m. Pooled results are two independent experiments