Journal: PLoS ONE

Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

doi: 10.1371/journal.pone.0052627

Figure Lengend Snippet: Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on fibronectin-coated substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

Techniques: Staining

Journal: PLoS ONE

Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

doi: 10.1371/journal.pone.0052627

Figure Lengend Snippet: Exo70 redistributes in Cav1-positive compartments upon cell detachment. ( A ) Hela cells expressing Cav1-mRFP and Exo70-GFP were kept in suspension for 1 h and replated on fibronectin for 3 h, and then visualized by confocal dual-colour spinning-disk microscopy (see corresponding movie S1). Arrow points to a dynamic Cav1- and Exo70-positive vesicle. Bottom panel shows selected frames from the time-lapse series corresponding to the boxed region in the upper panel. Time is given in second. ( B ) Hela cells expressing Cav1-GFP and cavin-1-mRFP were treated as in panel A. Inset shows higher magnification of region indicated by an arrow. Scale bars, 5 µm.

Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

Techniques: Expressing, Microscopy

Journal: PLoS ONE

Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

doi: 10.1371/journal.pone.0052627

Figure Lengend Snippet: Actin and microtubule cytoskeletons act at distinct steps of Cav1 trafficking. ( A, B ) Hela cells expressing Cav1-mRFP maintained in suspension for 1 h, were replated on fibronectin for 3 h in the presence of 10 µM nocodazole (panel A) or 10 µg/ml cytochalasin-B (panel B) for 30 min. Cells were then analyzed by time-lapse confocal spinning disk microscopy. The right panels represent selected frames from the time-lapse series (time is given in second). Arrows point to Cav1-positive intracellular vesicles. See corresponding movie S2 (panel A) and movie 3 (panel B). Scale bars, 5 µm.

Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

Techniques: Activated Clotting Time Assay, Expressing, Microscopy

Journal: PLoS ONE

Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

doi: 10.1371/journal.pone.0052627

Figure Lengend Snippet: Exo70 is required for Cav1 transport to the plasma membrane. ( A, B ) Hela cells expressing Cav1-mRFP and α5-integrin-GFP either mock-treated (A) or treated with a specific siRNA to silencing Exo70 (B) were maintained in suspension for 1 h and replated on fibronectin for 3 h, and visualized using time-lapse spinning disk microscopy. Scale bars, 5 µm. ( A′, B′ ) Intensity profile of Cav1 (red) and α5-integrin (green) along the white lines shown in panel A and B. ( C ) Co-localization analysis of Cav1-mRFP and α5-integrin-GFP in cells as in panels A and B in a 20-pixel width region along the cell periphery of mock- or Exo70 siRNA-depleted cells using two specific siRNAs (siRNA 7c and 7d). Results are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

Techniques: Expressing, Microscopy

Journal: Stem Cells (Dayton, Ohio)

Article Title: Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial‐Like Cells

doi: 10.1002/stem.2750

Figure Lengend Snippet: SB431542 exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p

Article Snippet: SB431542 (10 µM) was added for 1, 2, and 3 days to the differentiation media in combination with BMP4, RA, and EGF as detailed in Figure A. qRT‐PCR analysis at day 20 indicated the highest expression of ΔNp63 in groups treated for 2 and 3 days with SB431542 (Fig. B).

Techniques: Real-time Polymerase Chain Reaction

Journal: Biophysical Journal

Article Title: Simulations Reveal Multiple Intermediates in the Unzipping Mechanism of Neuronal SNARE Complex

doi: 10.1016/j.bpj.2018.08.043

Figure Lengend Snippet: Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), syntaxin (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain

Article Snippet: Synaptobrevin (Sb) and syntaxin (Sx) both contribute to this complex with one α -helix, whereas SNAP25 consists of two α -helical motifs connected by an unstructured linker loop.

Techniques: