Journal: Traffic (Copenhagen, Denmark)

Article Title: Spastin Couples Microtubule Severing to Membrane Traffic in Completion of Cytokinesis and Secretion

doi: 10.1111/j.1600-0854.2008.00847.x

Figure Lengend Snippet: A–F) In HeLa cells fixed with methanol, endogenous spastin (A and D) colocalizes with the midbody markers Aurora B (B) and PRC1 (E). Note the double-ring appearance of spastin on either side of the stembody, most obvious in (A). G–I) Spastin (G) also colocalizes with GFP-VPS4A(E235Q) (H) in double-ring structures in the midbody, in methanol-fixed HeLa cells.

Article Snippet: Rabbit polyclonal anti-GFP (6556), rabbit polyclonal anti-aurora B and rat polyclonal anti-tyrosinated tubulin (YL1/2) were obtained from Abcam.

Techniques:

Journal: Traffic (Copenhagen, Denmark)

Article Title: Spastin Couples Microtubule Severing to Membrane Traffic in Completion of Cytokinesis and Secretion

doi: 10.1111/j.1600-0854.2008.00847.x

Figure Lengend Snippet: A–C) Hela (A and B) and MRC5 (C) cells were labeled with alpha-tubulin following spastin depletion with pooled siRNA oligonucleotides 1–4. Note long intercellular bridges (arrowheads) that were sometimes very convoluted (B). Alpha-tubulin-labeled puncta were often seen in association with these bridges (arrow in B). D and E) The intercellular bridges (arrowheads) were also seen following spastin depletion using two individual spastin siRNA oligonucleotides. Successful spastin depletion in these experiments is verified in (F). G–I) The intercellular bridges (arrowheads) typically joined two cells, as shown in DIC image (G) of YFP–tubulin (H)-expressing HeLa cells depleted of spastin. J–L) Some of the intercellular bridges had the appearance of very elongated midbodies, which labeled with midbody markers [e.g. aurora B; (K)] as well as with MT markers (J). M–O) Hela cells transfected with 60 kD myc-spastinK388R (M) and labeled for alpha-tubulin (N) also displayed similar intercellular bridges (arrowhead). Formaldehyde was used in fixed preparations except (J–L) where methanol was used.

Article Snippet: Rabbit polyclonal anti-GFP (6556), rabbit polyclonal anti-aurora B and rat polyclonal anti-tyrosinated tubulin (YL1/2) were obtained from Abcam.

Techniques: Labeling, Expressing, Transfection