Supplementary Figure 3: Characterization of dynein-binding motifs of Spindly. Related to Figure 1.
(a) Immunoblot showing Spindly knockdown and expression of siRNA-resistant GFP-tagged versions of Spindly in mitotic population. The experiment was repeated twice with similar results. (b) Representative immunofluorescence images of HeLa cells transfected with siRNA to Spindly and treated or not with doxycycline (Dox) to induce the expression of GFP-Spindly variants, and immunostained for the indicated antigens. The experiment was repeated at least three times with similar results. The same images of FL and SpindlyΔN are shown in Figure 1f. (c) Representative images of nocodazole-treated HeLa cells transfected with siRNAs to Spindly and expressing the indicated GFP-Spindly variants, and immunostained for the indicated antigens. Quantification is shown in Figure 1c. (d) Representative images of metaphase HeLa cells transfected with siRNA to Spindly and expressing the indicated GFP-Spindly variants, and immunostained for the indicated antigens. Quantification is shown in Figure 1e. The same images of FL, SpindlyΔSB and SpindlyΔCCS are shown in Figure 1f. FL, Full length Spindly.
