M1 Spastin’s Hp motif directly inserts into the lipid monolayer of LDs. (A) Diagrams of synthetic constructs. Amino acid number and protein domains are indicated as in Fig. 1 A. (B) Localization of mEmerald-Sec61β and mApple-M1^1–92 monitored by SIM in HeLa cells treated with 300 µM oleic acid for 16 h. Representative images and relative intensity profile are shown. (C) Localization of mEmerald-Sec61β and mApple-M1 Spastin monitored by SIM in HeLa cells treated with oleic acid. Representative images and relative intensity profile are shown. (D) Changes in intensity of 2xFKBP-mApple-M1^1–92 in BODIPY-493/503–labeled LDs following 1 µM AP20187 treatment in HeLa cells. Representative confocal images are shown. (E) Changes in relative intensity of 2xFKBP-mApple-M1^1–92 as described in D. Means ± SEM are shown (12 cells from two independent experiments). (F) DsRed2-M1^1–92 colocalizes with LDs in HeLa cells in steady state. Representative confocal images are shown.