Mutation N154A increases the resistance of actin waves to latrunculin.

<p>Cells expressing mRFP-lifeact only or co-expressing mRFP-lifeact and either GFP-MIB or GFP-tagged MIB mutants were starved for 1 h at which time 4.2 µM latrunculin was added. Cells were scored for the presence of actin waves by mRFP-lifeact fluorescence as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094306#pone-0094306-g003" target="_blank">Fig. 3</a>. Short movies (10 frames every 10 s) of the fields of cells were taken before and after adding latrunculin and the percent of cells with waves was calculated. (A) <i>myoB⁻</i> -cells co-expressing lifeact and MIB (open circles) or lifeact and N154A (black circles). Time = 0 corresponds to cells before addition of latrunculin. (B) <i>myoB⁻</i>-cells expressing lifact alone (black circles), parent AX2 cells expressing lifeact alone (grey circles) and <i>myoB⁻</i>-cells co-expressing lifeact and MIB (open circles). Time = 0 corresponds to cells before addition of latrunculin. (C) <i>myoB⁻</i>-cells expressing lifeact only or co-expressing lifeact and either Tail, N154A-BH-Ala or dGPQSH3 in the absence (grey bars) and presence (black bars) of latrunculin. Movies were recorded before addition of latrunculin (No Latr) and between 60 and 80 min after addition of latrunculin (+Latr) at which time the percentage of cells with waves plateaus. The results are representative of at least 2 independent experiments. Error bars were calculated using the binomial probability confidence interval calculator developed by Daniel S. Soper (<a href="http://www.danielsoper.com/statcalc" target="_blank">http://www.danielsoper.com/statcalc</a>) and correspond to the 95% confidence interval.