Fig. 4

Influence of AH charge on the selective targeting of Plin4 to LDs. a Helical wheels depicting the mutations that were introduced into Plin4 33mer. In this series, charged residues in the polar face were mutated, and the hydrophobic face was either kept intact or modified with the previously characterized 2T→V mutation (Fig. 3). Charge-swap mutant is abbreviated as ‘csw’. The plot shows the hydrophobicity and hydrophobic moment of each mutant as calculated using Heliquest⁷¹. b Representative images of the mutants expressed as mCherry fusions in HeLa cells LDs were stained with Bodipy. All mutants were prepared as identical 4-mer repeats. Scale bar: 10 µm. c Quantification of HeLa images, showing % of LDs per cell positive for the indicated Plin4 construct. 30–40 cells per experiment were quantified for each construct and the error bars depict the range of means between two independent experiments. Two sets of mutants were analyzed at different times in the project and are therefore presented on separate plots. d Representative images of the mutants expressed as GFP fusions in budding yeast. LDs were marked with Erg6-RFP. For consistency, the colors of the yeast images are inverted. Scale bar: 5 µm