Figure 1. A high-throughput protoplast-based assay enables examination of thermoresponsive condensate formation.

(a) Bar graph showing the top 20 enriched GO category (molecular function) of predicted PLCDs with fold enrichment shown on x-axis and GO categories on y-axis. Plot was generated using ShinyGO (https://bioinformatics.sdstate.edu/go/). (b) Violin plot representing the daily temperature fluctuation in Columbia, Missouri through the year 2024. Orange color (high temperature) and cyan color (low temperature). The highlighted rectangular area represents the optimum growing season of Arabidopsis thaliana. Data collected from https://weather.com. (c) Arabidopsis mesophyll cells were isolated by enzymatic cell wall digestion, allowing transfection with protein expression plasmids. Protoplasts were then incubated under cold (4 °C), normal (22 °C), or heat (37 °C) conditions for 30 minutes. (d) The system recapitulates known stress granule dynamics, as demonstrated by the condensation behavior of the stress granule protein RBP45B. The images show fluorescent signal (YFP) and the corresponding bright field images taken at 4 °C (left), 22 °C (middle) and 37 °C (right). Scale bar = 10 μm.