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. 2011 Dec 12;195(6):953–963. doi: 10.1083/jcb.201104142

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© 2011 Gong et al.

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Figure 4. — Fsp27 promotes LD growth by mediating directional lipid transfer. (A) Time-lapse images showing lipid transfer in 3T3-L1 preadipocytes expressing Fsp27-GFP. The red asterisks and yellow arrowheads represent the acceptor and donor LDs, respectively. (B–D) Size comparison of the donor and acceptor LDs of active (20 pairs) or inactive (15 pairs) lipid transfer pairs. Line arrows indicate contacted LD pairs. NS, no significant difference. ***, P < 0.001. Error bars in D represent SEM. (E) Based on data in D, the percentage of LD pairs with different diameter ratios that underwent active lipid transfer was calculated from 35 LD pairs out of 20 independent videos. (F) Time-lapse DIC images showing lipid transfer in 3T3-L1 preadipocytes expressing Fsp27-GFP and untagged Fsp27 (coexpressed with GFP or GFP-Plin). GFP images were taken in three or five key time frames. Arrowheads and asterisks represent the donor and acceptor contacted LD pair, respectively. Error bars represent SD. (G) The fate of Plin on smaller LDs during Fsp27-mediated lipid transfer. Cherry-Plin and Fsp27-GFP were coexpressed in 3T3-L1 preadipocytes. Cherry-Plin signal on the large LD was photobleached (boxed regions). (H) Fsp27 promotes lipid transfer and LD growth in vitro. Size distribution of LDs isolated from 293T cells expressing GFP, Fsp27-GFP, or Fsp27-KKRA-GFP before and after 1 h of incubation in vitro. Each distribution was calculated from 120–150 LDs. The experiment was repeated for three times, and a representative one is shown here. (I) Images of contacted LDs with Fsp27-GFP enrichment at the LDCS before and after incubation. Bars: (A) 4 µm; (F and I) 5 µm; (G) 2 µm.