Figure 4. The Hyd and the Pos-Seq cooperate for targeting to LDs.

A) Schematic representation of the mutants used in this figure. The three studied sequences have been colored: Hyd (red), the Jux (blue) and the End (orange). Numbers on the residues indicate the relative position of each residue within the full-length protein with respect to the N-terminal end (see in addition Table 1). Letters between brackets indicate the corresponding panel for each mutant. Mutants were N-terminally tagged with GFP, expressed in lipid-loaded cells and their final distribution was analyzed by fluorescence microscopy (B–H). Arrows mark the selected areas for high magnification (G and H). Shows the colocalization between the Hyd (green) and endogenous Sec61 (red) and the Hyd-End³ (green) with Nile Red (red). I) Distribution of the mutants in sucrose density gradients as described in Figure 1. J and K) Low magnification panels to illustrate that cells transfected as above with Hyd and with ER (in green) display a normal formation of LDs (in red), although the mutants accumulate in the ER or the cytosol, respectively. Discontinuous lines indicate the edge of transfected cells. Scale bar is 20 μm.