Figure 6.

Liposome-bound LC3 stimulates JMY’s nucleation activity. (A) Liposome-bound LC3 promotes JMY’s intrinsic nucleation activity more potently than an equal amount of soluble JMY in a pyrene-actin polymerization assay. Time courses of actin assembly with liposome-bound LC3 (5.25 µM) are shown in three replicas (yellow, cyan, and pink) to confirm no error in pipetting of liposomes. (B) Dose-dependence of liposome-bound LC3 enhancement of JMY’s nucleation activity (middle). The initial slope (first 250 s) of each curve is normalized and plotted as a proxy for nucleation rate (right). Pyrene-actin polymerization assays were performed at 23°C in 50 mM KCl, 1 mM MgCl₂, 1 mM EGTA, and 10 mM Imidazole, pH 7.0, with 1 µM actin, 200 nM JMY, 5.25 µM soluble LC3, and liposome-bound LC3 as indicated.