Figure 8.
FIT2-interacting proteins at the nascent LD formation site. (A) COS-7 cells expressing FIT2-mCherry and LiveDrop were delipidated for 72 h and then treated with 0.2 mM OA (final concentration) to induce nascent LD formation. Live cell images were acquired by GI-SIM after OA addition at t = 0. Magenta arrowheads indicate the accumulation of FIT2, and green arrows indicate the nascent LDs. Scale bar, 1 µm. (B) As in A, but with cells expressing REEP5-mCherry and LiveDrop. Magenta arrows indicate the accumulation of REEP5, and green arrows indicate the nascent LDs. Scale bar, 1 µm. (C) Representative 3D-SIM images of colocalization between LiveDrop and endogenous septin 2 in delipidated COS-7 cells with or without OA treatment. Enlargements of the boxed regions are shown. Scale bar, 5 µm; 1 µm (inset). (D and E) As in C, but in COS-7 cells transfected with siControl and siSeipin. The Mander’s coefficient was measured by ImageJ. In each group, 45–50 ROIs in 10–15 cells were measured. Unpaired t test, ***, P < 0.001. Scale bar, 5 µm; 1 µm (inset). Error bars represent SEM. (F) A model of FIT2-mediated nascent LD formation. ROI, region of interest; siControl, small interfering Control; siFIT2, small interfering FIT2; siSeipin, small interfering seipin; Starv., starvation.

FIT2-interacting proteins at the nascent LD formation site. (A) COS-7 cells expressing FIT2-mCherry and LiveDrop were delipidated for 72 h and then treated with 0.2 mM OA (final concentration) to induce nascent LD formation. Live cell images were acquired by GI-SIM after OA addition at t = 0. Magenta arrowheads indicate the accumulation of FIT2, and green arrows indicate the nascent LDs. Scale bar, 1 µm. (B) As in A, but with cells expressing REEP5-mCherry and LiveDrop. Magenta arrows indicate the accumulation of REEP5, and green arrows indicate the nascent LDs. Scale bar, 1 µm. (C) Representative 3D-SIM images of colocalization between LiveDrop and endogenous septin 2 in delipidated COS-7 cells with or without OA treatment. Enlargements of the boxed regions are shown. Scale bar, 5 µm; 1 µm (inset). (D and E) As in C, but in COS-7 cells transfected with siControl and siSeipin. The Mander’s coefficient was measured by ImageJ. In each group, 45–50 ROIs in 10–15 cells were measured. Unpaired t test, ***, P < 0.001. Scale bar, 5 µm; 1 µm (inset). Error bars represent SEM. (F) A model of FIT2-mediated nascent LD formation. ROI, region of interest; siControl, small interfering Control; siFIT2, small interfering FIT2; siSeipin, small interfering seipin; Starv., starvation.

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