Fig. 4: The interplay with chaperones and co-chaperones is mediated by the double α-helix of the PLD.

A Turbidity measurements of the PLD in presence of the indicated chaperones (all in 1:2 molar ratios). Error bars are not included in the PLD plot (black dots) for clarity. B DIC microscopy image of the condensates formed by 20 μM PLD in complex with HSP72 after 48 h of incubation at 25 °C. C NMR signal intensity plots of 25–35 μM PLD in complex with the indicated chaperones (all in 1:2 molar ratios). For comparison, the plots are overlaid with the data corresponding to de-mixed PLD (300 μM PLD, broken purple line, Fig. 1F) and HSP72 interaction (orange broken line). D NMR intensity plots for MetO PLD in the presence of the chaperones (green bars) in comparison to the unmodified PLD:chaperone interactions in identical molar ratios (broken lines). E LLPS of 20 μM PLD in the presence of HSP72 and/or JDPs represented as the area under the curve of the turbidity measurements after 24 h. Vertical broken lines separate the three JDP classes (A, B, and C, indicated on top). F DIC microscopy image of the condensates formed by 20 μM PLD in complex with HSP72:DNAJB1 after 3 h of incubation at 25 °C. G NMR signal intensity plots for the interaction of 25–35 μM PLD with JDPs (all in 1:2 molar ratios). In each plot, the broken line represents the interaction of the PLD with HSP72 and the corresponding JDPs (all in 1:2:2 molar ratios). H LLPS of 20 μM PLD in presence of HSP90 and the specified co-chaperones as measured by turbidity. I LLPS of 20 μM MetO PLD in the presence of the specified chaperones and co-chaperones. For simplicity, the plots in C, D, and G show the reverse of the NMR signal decay. The gray broken line in E, H, and I represents the average turbidity of 20 μM PLD, for comparison. Scale bars (B, F) correspond to 25 μM. Turbidimetry and microscopy samples (A, B, E, F, H, I) contained 150 mM KCl, whereas NMR samples (C, D, G) contained 10 mM KCl.