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Fig. S1 Phylogenetic analysis of villins from angiosperms.

Fig. S2 Multiple sequence alignment of plant villin proteins.

Fig. S3 AtVLN4 severs or depolymerizes actin filaments in a calcium (Ca2+)-dependent manner.

Fig. S4 AtVLN4 inhibits actin filament elongation from barbed ends in a calcium (Ca2+)-dependent manner.

Fig. S5 Subcellular localization of GFP, AtVLN4-GFP and GFP-fABD2 in tobacco pollen.

Fig. S6AtVLN4 is abundant in the Arabidopsis root hair zone.

Fig. S7 Molecular analysis of AtVLN4 T-DNA insertion lines.

Fig. S8 Comparison of primary root length (mean ± SE, > 20) in wild-type and atvln4 plants.

Video S1 Time-lapse fluorescence microscopy series of 200 μM Oregon Green-actin filaments treated with fluorescence buffer only.

Video S2 Actin filaments severed by 0.5 nM AtVLN4 in the presence of 200 μM calcium (Ca2+).

Video S3 Actin filaments severed by 1 nM AtVLN4 in the presence of 200 μM calcium (Ca2+).

Video S4 Actin filaments severed by 2.5 nM AtVLN4 in the presence of 200 μM calcium (Ca2+).

Video S5 Actin filaments severed by 5 nM AtVLN4 in the presence of 200 μM calcium (Ca2+).

Video S6 Actin filaments show minimal breakage when treated with 5 nM AtVLN4 in the absence of calcium (Ca2+).

Video S7 Actin filaments show minimal breakage when treated with 5 nM AtVLN4 in the presence of 0.5 μM calcium (Ca2+).

Video S8 Actin filaments severed by 5 nM AtVLN4 in the presence of 5 μM calcium (Ca2+).

Video S9 Actin filaments severed by 5 nM AtVLN4 in the presence of 50 μM calcium (Ca2+).

Video S10 This movie corresponds to the pattern of cytoplasmic streaming of a wild-type root hair (shown in Fig. 6a, left panels).

Video S11 This movie corresponds to the pattern of cytoplasmic streaming of an atvln4-1 root hair (shown in Fig. 6a, middle panels).

Video S12 This movie corresponds to the pattern of cytoplasmic streaming of an atvln4-2 root hair (shown in Fig. 6a, right panels).

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