Figure EV4. C99's cholesterol‐binding domain facilitates MAM formation and activation.

• A
  Quantification of MAM activity by measurement o f phospholipid synthesis and transfer between ER and mitochondria in the indicated cells (Montesinos et al, 2020). Graphs represent the levels of ³H‐serine incorporation into phosphatidylserine (PS) and its subsequent conversion into phosphatidylethanolamine (PE) and phosphatidylcholine (PC) over the indicated time periods. Two‐way ANOVA (Time, Group) (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001).
• B
  Cholesterol efflux, measured 4 h after completion of 1 h pulse‐chase with ³H‐cholesterol was assayed in APP‐DKO or WT (APP‐WT) cells transfected with an empty vector (EV) or the C99^WT or C99^MUT constructs. One‐way ANOVA (n = 3; *P < 0.05, ***P < 0.001).
• C
  Quantification of neutral and acidic SMase activities in the indicated conditions. One‐way ANOVA (n = 3; *P < 0.05).
• D
  Representative images of APP‐DKO cells expressing C99^WT‐GFP or C99^MUT‐GFP and stained with LipidTox (in red) to detect lipid droplets (white arrows). Scale bar = 15 μm. Graph on the right shows the proportion of cells with lipid droplets. EV, empty vector. One‐way ANOVA (n = 3; *P < 0.05).

Source data are available online for this figure.