Figure 2. PLCD-containing proteins exhibit variability in thermoresponsive behavior.

(a) k-means clustering with silhouette analysis identifies four distinct clusters of condensation behavior across 4 °C, 22 °C, and 37 °C. Proteins were assigned to the nearest cluster centroid based on Euclidean distance. Cluster 1 proteins display high condensation at all temperatures. Cluster 2 proteins show low condensation at 4 °C and 22 °C and moderate condensation at 37 °C. Cluster 3 proteins exhibit moderate condensation at 4 °C and 22 °C and high condensation at 37 °C. Cluster 4 proteins show low condensation at 4 °C and 22 °C and high condensation at 37 °C. (b) Percentage of cells displaying mVenus-AtProtein puncta at each tested temperature. For each temperature, in average 50 cells were examined at each temperature. The protoplast count from two replicates were combined to plot the graph. (c) Representative protoplast images from each cluster at 4 °C, 22 °C, and 37 °C. FLL2 from Cluster 1, bZIP10 from Cluster 2, FH7 from Cluster 3 and LARP1C from Cluster 4. The images show the YFP signal (Left) and bright filed image (right) of each protein. Scale bar = 10 μm. N indicates the nucleus.