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. 2018 Mar 16;59(5):805–819. doi: 10.1194/jlr.M081679

Fig. 5.

Fig. 5.

Stx17 regulates the redistribution of ACSL3 to nascent LDs. A: HeLa cells transiently expressing GFP-KDEL were mock-transfected or transfected with siRNA Stx17 (440), starved for 24 h in the absence of FCS followed by incubation with OA for 1 h, fixed, and then stained with LipidTox. Bar, 5 μm. The degree of colocalization between LipidTox and GFP-KDEL was measured with Manders’ coefficients. NS, not significant. B: HeLa cells were mock-transfected or transfected with siRNA Stx17 (440) or siRNA targeting ACSL3, and incubated for 72 h without FCS for the last 24 h. OA was added, and cells were incubated for 1 h, fixed, stained with LipidTox and an antibody against ACSL3, and quantified. Values are the means ± SD (n = 3). C: HeLa cells were treated as described in A and stained with LipidTox and an antibody against ACSL3 or Tip47. Bars, 200 nm. D: Quantitation of data in C. *P ≤ 0.05; NS, not significant. The fluorescence intensities of ACSL3 and Tip47 surrounding LDs relative to that of LipidTox were plotted. Values are the means ± SEM (n = 3). E: Nascent LDs were analyzed by FRAP using a protocol depicted in the figure.