(A) Helical wheel representation of the AHs of Plin1 (aa 110–189 aa), Plin2 (aa 101–191 aa), Plin3 (aa 114–204), and Plin4 (aa 246–377, corresponding to the Plin4 4mer construct). In the case of Plin1, Plin2, and Plin3, the predicted AH regions are interspersed by short aa linkers, which are also indicated. Diagrams above the helical wheels show the full-length proteins, with AH regions shown in orange and the four-helix bundle in dark gray. (B) AA composition of the AH of Plin1, 2, 3, and 4 (in %) in comparison with the average aa composition of vertebrate proteins (av. vert). The blue and red backgrounds indicate lower or higher % as compared to vertebrate values, respectively. (C) Localization of GFP fusions with the AH region of Plin1, Plin2, Plin3, or Plin4 in S. cerevisiae cells. The experiment was performed with wild-type yeast cells (upper row), with pet10Δ cells (medium row) grown for 24 hr to stationary phase, or with pet10Δ cells grown to stationary phase and then transferred for 24 hr to oleic acid (OA) medium (lower row). Scale bar: 5 µm. (D) Bar plots of the percentage of cells showing intracellular puncta for the different proteins expressed. Sixty cells per each condition were counted in one of at least two representative experiments. (E) Quantification of the size distribution of fluorescent LDs (labeled with GFP-fusion proteins) in pet10Δ + OA cells. The plots show representative measurements from two independent experiments, where the following number of LDs was counted: Plin1 AH, 141; Plin2 AH, 143; Plin3 AH, 136; Plin4 6mer, 148; Plin4 12mer, 159. Pixel size: 0.091 μm x 0.091 μm.