(A) Expression of two distinct Pat1 binding mutants, Dhh1R295D and Dhh1S292DN294D, leads to a drastic reduction in PB formation. Cells co-expressing the indicated PB components in either Dhh1, Dhh1R295D or Dhh1S292DN294D cells were grown in SCD media to exponential growth phase, then shifted to glucose-rich or glucose-starvation conditions for 30 min and observed by fluorescence microscopy. Scale bar: 5 µm. (B) Quantification of images in A depicting number of Dhh1 positive and Dcp2 positive PBs/cell; N = 4 biological replicates with >300 cells/replicate, SEM. (C) Dhh1R295D and Dhh1S292DN294D mutants are defective in PB formation upon hippuristanol treatment. Cells expressing either Dhh1, Dhh1R295D, Dhh1S292DN294D or dhh1Δ, were grown in SCD media to exponential growth phase after which they were treated with 10 µM hippuristanol for 3 hrs. The kinetics of PB formation was monitored using Dhh1-GFP and its co-localization with Dcp2-mCherry. (D) Quantification of PBs/cell. Stills from the live imaging time course are shown. N = 3 biological replicates with >300 cells/replicate, SEM.