Journal: bioRxiv

Article Title: Cell intrinsic mechanical regulation of plasma membrane accumulation in the cytokinetic furrow

doi: 10.1101/2023.11.13.566882

Figure Lengend Snippet: (A) Representative images of mitotic and cytokinetic L1210 cells expressing H2B-GFP (blue, DNA) and labeled for cell surface protein content (orange/yellow). Cells were imaged immediately after labeling. Each image is a separate cell. (B) Quantifications of cell surface protein content as a function of distance from the division plane, as indicated in the schematic on top. Lines and shaded areas depict mean±SD, n depicts the number of individual cells in each cell cycle stage. (C) Percentage of cytokinetic cells that exhibit the accumulation of plasma membrane proteins at the cleavage furrow in the indicated cell lines. Data depicts mean±SD of independent experiments. N depicts the number of independent experiments and n depicts the total number of cytokinetic cells. (D, E) Representative images of cytokinetic BaF3 (D) and Hela (E) cells expressing H2B-GFP (blue) and labeled for cell surface protein content (orange/yellow). Scale bars denote 10 μm.

Article Snippet: The L1210 cells were obtained from ATCC (#CCL-219), the BaF3 cells were obtained from RIKEN BioResource Center, and the HeLa cells were gifted by the laboratory of Matthew Vander Heiden at Massachusetts Institute of Technology.

Techniques: Expressing, Labeling, Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Cell intrinsic mechanical regulation of plasma membrane accumulation in the cytokinetic furrow

doi: 10.1101/2023.11.13.566882

Figure Lengend Snippet: (A) Two competing models that can explain the accumulation of plasma membrane proteins at the cytokinetic furrow. (B-D) Representative images of mitotic and cytokinetic L1210 cells expressing H2B-GFP (blue, DNA) and labeled for plasma membrane (purple/white) using CellMask (panel B, N=3 independent experiments, n=46 cytokinetic cells), WGA (panel C, N=2 independent experiments, n=31 cytokinetic cells), or Filipin III (panel D, N=3 independent experiments, n=68 cytokinetic cells). Scale bars denote 10 μm. (E) Representative SEM images of fixed L1210 cells in cytokinesis. N=3 independent experiments, n=45 cytokinetic cells. Scale bars denote 2 μm.

Article Snippet: The L1210 cells were obtained from ATCC (#CCL-219), the BaF3 cells were obtained from RIKEN BioResource Center, and the HeLa cells were gifted by the laboratory of Matthew Vander Heiden at Massachusetts Institute of Technology.

Techniques: Clinical Proteomics, Membrane, Expressing, Labeling

Journal: bioRxiv

Article Title: Cell intrinsic mechanical regulation of plasma membrane accumulation in the cytokinetic furrow

doi: 10.1101/2023.11.13.566882

Figure Lengend Snippet: (A) Three mutually non-exclusive biophysical processes that can explain the plasma membrane accumulation at the cytokinetic furrow. (B) The experimental setup: Pre-labelled plasma membrane proteins are imaged over time to examine whether the pre-labelled proteins accumulate at the cytokinetic furrow, indicative of membrane movement on the cell surface. Since internal membranes are not labelled, exocytosis-driven membrane accumulation will not result in a detectable signal accumulation. (C) Representative timelapse imaging of L1210 cells expressing H2B-GFP and labeled for cell surface protein content (orange/yellow). N=3 independent experiments, n=14 cytokinetic cells for L1210 cells. N=4 independent experiments, n=15 cytokinetic cells for HeLa cells. Scale bars denote 10 μm. (D) Quantifications of L1210 cell surface protein content at the cytokinetic furrow (orange) and cell poles (blue) as a function of time from anaphase onset. Lines and shaded areas depict mean±SD (n=14 cells). Data is normalized to the average of values at t≤0.

Article Snippet: The L1210 cells were obtained from ATCC (#CCL-219), the BaF3 cells were obtained from RIKEN BioResource Center, and the HeLa cells were gifted by the laboratory of Matthew Vander Heiden at Massachusetts Institute of Technology.

Techniques: Clinical Proteomics, Membrane, Imaging, Expressing, Labeling

Journal: bioRxiv

Article Title: Cell intrinsic mechanical regulation of plasma membrane accumulation in the cytokinetic furrow

doi: 10.1101/2023.11.13.566882

Figure Lengend Snippet: (A) Predicted plasma membrane accumulation at the furrow as furrow ingression progresses for different values of the membrane-cortex drag coefficient , membrane lipid-protein drag coefficient , and cortical contractility (B) Comparison of plasma membrane accumulation at the cytokinetic furrow in the force balance model and in live cell timelapse imaging as furrow ingression progresses. Lines depict modeling results using indicated model parameter values. Each orange dot depicts a cell at a given moment during furrow ingression (n=14 cytokinetic cells with a total of 103 images at different stages during furrow ingression). (C) Relative contribution of apparent cell surface area changes, in comparison to cortex driven membrane flows, to the accumulation of plasma membrane at the cleavage furrow as furrow ingression progresses. (D) Representative images of anaphase L1210 cells expressing H2B-GFP (blue) and labeled for surface protein content (orange/yellow). The cells were treated with DMSO (n=42 anaphase cells), 50 μM (-)-Blebbistatin (n=24 anaphase cells), or 2 μM Latrunculin B (n=35 anaphase cells) for 2 hours. Scale bars denote 10 μm.

Article Snippet: The L1210 cells were obtained from ATCC (#CCL-219), the BaF3 cells were obtained from RIKEN BioResource Center, and the HeLa cells were gifted by the laboratory of Matthew Vander Heiden at Massachusetts Institute of Technology.

Techniques: Clinical Proteomics, Membrane, Comparison, Imaging, Expressing, Labeling