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. Author manuscript; available in PMC: 2024 Mar 15.
Published in final edited form as: Cell Rep. 2024 Feb 13;43(2):113761. doi: 10.1016/j.celrep.2024.113761

Figure 3. FAD PSEN1 mutations stabilize γ-Secretase E-S complexes.

Figure 3.

(A) Design of fluorescence lifetime imaging microscopy (FLIM) experiments.

(B) Microscopic imaging with pseudo-color analysis to identify C99 or Aβ intermediate-rich subcellular areas (regions of interest [ROIs], circles), followed by FLIM, in HEK293 cells.

(C and D) Quantification of FLIM results reveals that FAD mutations in (C) PSEN1 and (D) C99 increase stabilization of enzyme-C99 (left) or enzyme-Aβ intermediate complexes (right) in intact cells. Scale bars, 10 μm in upper micrographs and 5 μm in lower (inset) micrographs. For all graphs, n = 136–141 ROIs from 7 to 10 cells; one-way ANOVA and Tukey’s multiple comparisons test; n.s., p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.