JCS013102 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. After mitosis, CAP350 is concentrated in numerous dotted areas where the Golgi complex reforms. HeLa cells during telophase co-stained with anti-CAP350 (left) and anti-CTR433 (right) antibodies with the latter recognising the Golgi. Note that CAP350-enriched dots around the centrosome are located where the Golgi reforms. Bar, 20 μm.

• Supplemental Figure S2 -

  Fig. S2. Overexpressed full-length Myc-tagged CAP350 localises to a subset of pericentrosomal microtubules. (A) Overexpression of full-length Myc-tagged CAP350 in cells expressing α-tubulin GFP. Full-length CAP350 localises on a subset of pericentrosomal MTs. (B) 4.5× magnification of the boxed area in A. Bar, 20 μm.

• Supplemental Figure S3 -

  Fig. S3. Effect of CAP350 depletion on the microtubule network. Cells were transfected with the GL2 (A) or CAP350 (B) duplexes for 48 hours and stained for CAP350 and tubulin. CAP350 depletion has no obvious effect on the whole MT network organisation. Bar, 20 μm.

• Supplemental Figure S4 -

  Fig. S4. Microtubule and CAP350 organisation in polarised MDCK cells. (A) Apical view of a 3D reconstruction of a group of polarised MDCK cells labelled with CAP350 (green) and DAPI (blue). CAP350 is concentrated at the centrosomes and diffuse CAP350 dots are dispersed in the cytoplasm. Note that there is no evidence of CAP350 accumulating at the apical peripheral anchoring sites where the apico-basal microtubule minus ends are anchored. (B) Schematic diagram depicting microtubule (red lines) and CAP350 (green dots) organisation in a polarised MDCK cell. The majority of the microtubules are arranged in an apico-basal array (the reduced apical radial array and basal microtubule network are not shown). CAP350 is concentrated at the apical centrosome with a few cytoplasmic dots scattered throughout the apical region of the cell. The nucleus is labelled with DAPI (blue). (C) Lateral view of three of the polarised MDCK cells from A, showing CAP350 concentrated at the two centrioles and diffuse dots distributed within the apical cytoplasm. Bar, 5 μm.