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. 2012 Jun 21;7(6):e39774. doi: 10.1371/journal.pone.0039774

Figure 1. Actin depolymerization increases the amount of endosomal WASH.

Figure 1

(A) 3T3 cells were loaded with fluorescent Transferrin (Tf) until equilibrium, then treated with 0.2 µM Latrunculin A (LatA) for 10 min, 1 µM CytochalasinD (CytoD) for 30 min, or carrier in the continuous presence of Tf. Cells were processed for immunofluorescence using WASH antibody and observed by spinning disk confocal microscopy. A single plane is displayed. Scale bar: 10 µm (1 µm in inserts). LatA and CytoD treatments increase the fluorescence signal of WASH. (B) Since endosomes are clustered in the perinuclear region, the increase of WASH intensity was quantified on isolated endosomes from the cell periphery (see Methods). Both LatA and CytoD treatments induce a statistically significant increase of the WASH fluorescence signal (n refers to the number of endosomes, *: p<0.001, one way ANOVA followed by a Tukey pairwise comparison).