Fig. 5: Related MAP4Ks that interact with TOT3 play a role in regulating hypocotyl growth under warm temperature.

a Overlap between TOT3 interactome (based on tandem affinity purification (TAP) and immunoprecipitation (IP)) and differentially phosphorylated proteins in tot3-2 versus Col-0 (referred to as tot3 phosphoproteome obtained from 3-day-old tot3-2 and Col-0 seedlings grown at 28 °C). b Yeast two-hybrid validation of the interaction between TOT3 and TOI4/5. BD, BINDING DOMAIN; AD, ACTIVATION DOMAIN; -LT, without Leu and Trp; -LTH, without Leu, Trp and His. c TOI4/5 phosphosites significantly deregulated in tot3 phosphoproteome. Each dot represents a biological replicate. Statistical analyses using Student’s t-test with two-sample unequal variance and two-tailed distribution; p-value is indicated. d, e Hypocotyl length of 7-day-old tot3, tot3 toi4, tot3 toi5, toi4 toi5 and Col-0 plants at 21 and 28 °C in short-day conditions. Representative pictures (d) and hypocotyl length quantification (e). Scale bar, 5 mm. Box plots show median with Tukey-based whiskers and outliers. The number of individually measured seedlings (n) is indicated above the X-axis. Fold-change is indicated for each genotype; and letters indicate significant differences based on two-way ANOVA and Tukey’s test (p < 0.01) (e). The p-value for the interaction (genotype × temperature) is shown at the top.