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Figure S1. DCL is a MAP and stabilizes microtubules. Panel I: DCL expression in transfected COS-1 cells. (A-C) DCL overexpression in COS-1 cells. (D-F) DCL overexpressed in COS-1 cells treated with colchicine. Green represents DCL; red represents α-tubulin and yellow indicates DCL co-localization with α-tubulin, demonstrating its association with microtubules. Blue represent (nuclear) DNA. Arrows indicate DCL-associated microtubule bundles, which are resistant to colchicine treatment. Also note the clear association of DCL with a centrosome in A. Scale bar is 10 μm. Panel II. In vitro microtubules polymerization by DCL. Different concentrations of recombinant DCL protein were incubated with purified tubulin. Turbidity of the DCL/tubulin mixture was monitored at 340 nm for 30 min with Taxol as a positive and water as a negative control. The graph shown is a typical example from multiple experiments (N=4) with similar results. Figure S2. DCL knock-down does not affect centrosome structure. Spickled DCL immunostaining (A, D, G) is highly concentrated (A, D) around centrosomes, as indicated by anti-γ-tubulin staining (B, E and H). Effective knock-down of DCL (G) does not affect structure or shape of the centrosomes (I). Merged illustrations of DCL and α-tubulin staining are shown of non-treated cells (C), cells transfected with siDCL-1 (F) and cells transfected with siDCL-3 (I). Green = DCL, Red = γ-tubulin, Yellow=colocalization of DCL and γ-tubulin. Scale bar is 10 μm.

FilenameFormatSizeDescription
EJN_5318_sm_FigureS1.tif5395KSupporting info item
EJN_5318_sm_FigureS2.tif4242KSupporting info item

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