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Results: 5

1.
Figure 2.

Figure 2. From: Rho-dependent control of anillin behavior during cytokinesis.

Myosin II localization can occur independently of F-actin and anillin but stable furrow positioning requires anillin. (A and B) Frames from time-lapse sequences of cells expressing MRLCSqh-GFP progressing through anaphase. (A) Control cell expressing MRLCSqh-GFP. (B) Cell treated with LatA. A projection of five 2-μm sections is shown (Video 6, available at http://www.jcb.org/cgi/content/full/jcb.200709005/DC1). (C and D) Cells expressing anillin-GFP (green in merged) fixed during anaphase/telophase in the presence of 1 μg/ml LatA, labeled with a Ser21 phospho-MRLCSqh antibody (red in merged; maximum intensity projections of multiple 0.25-μm deconvolved sections). (E) Frames from a time-lapse sequence of a MRLCSqh-GFP cell attempting cytokinesis after 3 d of anillin RNAi (see Video 7). (F) Cell expressing MRLCSqh-GFP (green in merged), fixed during anaphase in the presence of LatA after anillin RNAi and labeled with an anillin antibody (red in merged; maximum intensity projection of multiple 0.25-μm deconvolved sections). (G) Frames from a time-lapse sequence of a cell expressing MRLCSqh-GFP attempting cytokinesis after 3 d of anillin RNAi and in the presence of LatA (projection of five 2-μm sections; see Video 8). Times are h:min:s from anaphase onset. Bars, 5 μm.

Gilles R.X. Hickson, et al. J Cell Biol. 2008 January 28;180(2):285-294.
2.
Figure 4.

Figure 4. From: Rho-dependent control of anillin behavior during cytokinesis.

Anillin-containing structures in LatA form independently of MTs but associate with MT ends at the plasma membrane. (A) Time-lapse sequence of a cell expressing anillin-GFP and mcherry-tubulin exiting mitosis in the presence of colchicine (16 h) and LatA (1 h) after 3 d of Mad2 RNAi (max intensity projections of deconvolved 2-μm sections). (B) A nontransfected S2 cell treated with LatA and labeled with antibodies to anillin (green in merged) and α-tubulin (red in merged; max intensity projection of multiple deconvolved 0.25-μm sections). (C and D) Frames from time-lapse sequences of cells coexpressing anillin-GFP and mcherry-tubulin progressing through anaphase in the presence of 1 μg/ml LatA (see Video 10, available at http://www.jcb.org/cgi/content/full/jcb.200709005/DC1). Although the anillin-GFP channel was contrasted identically for all time points in each time-lapse series, the cherry-tubulin channel in Cand D was contrasted for each time point individually to counter the effects of photobleaching. Times are h:min:s from anaphase onset. Bars, 2 μm.

Gilles R.X. Hickson, et al. J Cell Biol. 2008 January 28;180(2):285-294.
3.
Figure 3.

Figure 3. From: Rho-dependent control of anillin behavior during cytokinesis.

Anillin recruits septin to the cleavage furrow and to filamentous structures in LatA. (A–C) Nontransfected (A and B) and anillin-GFP–expressing (C) S2 cells fixed during anaphase/telophase and labeled with antibodies to septinPnut (middle; red in merged) and anillin (A and B, left; green in merged) and the DNA stain HOECHST (blue in merged; maximum intensity projections of deconvolved 0.25-μm sections). C' is a 90° rotation about the y axis of the projected stack within the boxed region in C. (D) Time-lapse sequence of a cell expressing anillin-GFP attempting cytokinesis in the presence of LatA after 6 d of septinPnut RNAi. (E–G) Nontransfected S2 cells fixed during anaphase/telophase and labeled with antibodies to septinPnut (middle; red in merged) and Dia (left; green in merged) and the DNA stain HOECHST (blue in merged; maximum intensity projections of deconvolved 0.25-μm sections). (E) Control cell. (F) Cell after 3 d of anillin RNAi. (G) Cell after 3 d of anillin RNAi and LatA treatment combined. Bars, 5 μm.

Gilles R.X. Hickson, et al. J Cell Biol. 2008 January 28;180(2):285-294.
4.
Figure 5.

Figure 5. From: Rho-dependent control of anillin behavior during cytokinesis.

A model for anillin regulation and function during cytokinesis. Pathways controlled by RhoGEFPbl, including well-documented inputs into F-actin and myosin II (black) and a novel, separate input (pink) via anillin and septins (A). Through equatorial stimulation (1), MTs spatially control Rho activation and the independent recruitment of Dia, Rok, and anillin to the equatorial cortex. F-actin and Rok/myosin II contribute to equatorial focusing of anillin (and other cortical components) via cortical flow (2), whereas Rho–anillin–Septin complexes dynamically link contractile elements within the furrow to the plasma membrane (PM) and to MT plus ends, thus preventing the lateral instability of the furrow seen upon anillin RNAi. (B) LatA may stabilize Rho–anillin–Septin complexes by preventing their normal F-actin–dependent disassembly (A, blue). Continued assembly with blocked disassembly gives rise to the filamentous structures in LatA, perhaps mimicking midbody biogenesis that normally accompanies local loss of F-actin at the close of furrowing.

Gilles R.X. Hickson, et al. J Cell Biol. 2008 January 28;180(2):285-294.
5.
Figure 1.

Figure 1. From: Rho-dependent control of anillin behavior during cytokinesis.

RhoGEFPbl controls anillin-GFP localization during anaphase via F-actin– and myosin II–dependent and –independent mechanisms. (A–H) Frames from time-lapse sequences of anillin-GFP cells progressing through anaphase/telophase (single Z sections, except for D, G, and H, which are projections of five sections). (A) A control cell showing the normal redistribution of anillin-GFP (see Video 1, available at http://www.jcb.org/cgi/content/full/jcb.200709005/DC1). (B) A cell after 3 d of RhoGEFPbl RNAi (see Video 2). (C) A cell after 1-μg/ml LatA treatment (surface Z section; see Video 3). (D) The same as C but the LatA was washed out at 10 min, soon after the formation of anillin-GFP structures (see Video 4). (E) A cell after 3 d of RhoGEFPbl RNAi and LatA combined. (F) A cell after 3 d of MRLCSqh RNAi (see Video 5). (G) A cell after MRLCSqh RNAi and LatA combined. (H) A cell after 4 d of Dia and Rok RNAi and LatA treatment combined. (I) A LatA-treated anillin-GFP cell fixed during anaphase/telophase and labeled with a Rho1 antibody (red in merged; max intensity projection). (A', B', and F') semiquantitative measurements of the mean anillin-GFP intensities at the equator (□) and poles (◊), expressed as arbitrary units (AU) relative to time 0 (metaphase/anaphase [M/A]) in control, Pbl, and Sqh RNAi cells, shown for three cells each (different colors represent different cells). Times are h:min:s from anaphase onset. Bars, 5 μm.

Gilles R.X. Hickson, et al. J Cell Biol. 2008 January 28;180(2):285-294.

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