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. Author manuscript; available in PMC: 2009 Jun 1.
Published in final edited form as: J Cell Sci. 2008 Nov 4;121(Pt 23):3867–3877. doi: 10.1242/jcs.040477

Figure 2.

Figure 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 Latrunculin A (Lat-A) was examined by a pelleting assay. CapZ-capped F-Ce-actin (5 μM actin, CapZ : actin = 1:100) was incubated with various combinations of TMD-1, CeTM, and UNC-60B in the presence of 10 μM Lat-A for 15 min. 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 means ± SD of three experiments. (C) Kinetic measurements of depolymerization of CapZ-capped actin filaments. CapZ-capped F-Ce-actin (CapZ : actin = 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.