Figure 3. PLCDs of proteins display varying degrees of sufficiency in thermoresponsive condensation, which correlates to the thermoresponsive variability of full-length proteins.
(a,b,c) Bar plots representing the percentage of cells displaying mVenus-AtProtein puncta, mVenus-AtProteinPLCD (just PLCD) puncta and mVenus-AtProteinΔPLCD (deleted PLCD) puncta at 4 °C, 22 °C, and 37 °C in three distinct categories of PLCD sufficiency: (a) Category A PLCDs are neither sufficient nor necessary: full-length proteins condense upon heat stress even when PLCDs are deleted, while isolated PLCDs fail to condense. (b) Category B PLCDs are both sufficient and necessary: isolated PLCDs robustly condense at elevated temperature, and deletion of these domains diminishes condensation in the full-length protein. (c) Category C PLCDs are necessary but not sufficient: isolated PLCDs alone do not condense, but their deletion strongly reduces condensation in the full-length protein. (d) k-means clustering with silhouette analysis on the full-length proteins shows that Categories B and C have higher thermoresponsive variability than Category A, indicating that PLCD sufficiency maps to higher variability in the full-length protein. Higher and lower variability was defined by k-means clustering on percentage of cells with condensates across 4°C, 22°C, and 37°C. Proteins were assigned to the nearest cluster centroid based on Euclidean distance. (e) All Category A proteins map to Clusters 1 and 3 (low variability), whereas nearly all Category B and C proteins map to Clusters 2 and 4 (high variability). Line thickness corresponds to number of proteins, as indicated in the legend.