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Figure 2


Fig. 2. TMD-1 inhibits UNC-60B (ADF/cofilin)-dependent depolymerization of actin filaments in vitro. (A,B) Depolymerization of CapZ-capped actin filaments in the presence of Lat-A was examined by a pelleting assay. CapZ-capped F-Ce-actin (5 µM actin, CapZ:actin ratio of 1:100) was incubated with various combinations of TMD-1, CeTM, and UNC-60B in the presence of 10 µM Lat-A for 15 minutes. The mixtures were ultracentrifuged and fractionated into supernatants (sup) and pellets (ppt) and analyzed by SDS-PAGE (A) and densitometric quantification of depolymerized actin (%) in the supernatants (B). Data are the mean ± s.d. of three experiments. (C) Kinetic measurements of depolymerization of CapZ-capped actin filaments. CapZ-capped F-Ce-actin (CapZ: actin ratio of 1:100; 10% labeled with pyrene) was diluted to 0.5 µM actin with 1 µM Lat-A in the presence of indicated combinations of UNC-60B, CeTM, and TMD-1. Depolymerization of actin was monitored by decrease in pyrene fluorescence.





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