Fig. 3.

N-terminal predicted TMD of DHRS3 is necessary and sufficient for targeting to LD. (A) Schematic diagram of DHRS3-GFP deletion constructs. The location of the two predicted TMDs (pTMD) is indicated. (B) Oleate-treated HEK cells expressing the indicated DHRS3 fusion constructs (A) were separated into LD, cytosol (C), and membrane (M) fractions. Representative immunoblots of these fractions probed with the indicated antibodies are shown. Blot shown is representative of at least two replicates for each construct. (C) Chaotrope-resistant integration of DHRS3(1–60) and DHRS3(1–28) fusions into LD. Purified LDs (input) were treated with alkaline carbonate and repurified on a second sucrose gradient and analyzed by immunoblotting, as in Fig. 1. Representative immunoblots (n = 3) with the indicated antibodies are shown.