Fig. 4.

Split-luciferase protein interaction of Arabidopsis CaM, CML13, and CML14 with single and paired IQ domains of myosin ATM1 and ATM2 in planta. N. benthamiana leaves were infiltrated with Agrobacterium harboring respective NLuc–prey (myosin VIII IQ domains) and CLuc–bait (CaM, CMLs) vectors, and tested for luciferase activity 4 d later, as described in the Materials and methods. (A) Split-luciferase data are expressed as a fold increase or decrease relative to the RLU signal (log₁₀ scale) observed using the negative control bait CML42 which was set to an RLU of 1. Boxes contain each data point for six technical replicates, means are shown by a horizontal bar, the colored region is the 95% confidence interval, and whiskers extend to maximum and minimum data points. Asterisks indicate a significantly higher signal versus CLuc–CML42 as a negative control bait (one-way ANOVA against CML42 with Sidak’s test for multiple comparisons, *P<0.05, **P<0.01, ***P<0.001). Data are representative of at least three independent experiments. RLU, relative light units. Range of residues from each myosin tested were: ATM1 (894–943), ATM1-IQ1 + 2 (848–905), ATM1-IQ1 (848–879), ATM1-IQ3 + 4 (890–943), ATM1-IQ3 (890–928), ATM1-IQ4 (916–943), ATM2 (878–967), ATM2-IQ1 + 2 (878–949), ATM2-IQ1 (878–911), ATM2-IQ2 (899–949), and ATM2-IQ3 (922–967).