dynasore monohydrate  (Santa Cruz Biotechnology)

Journal: Molecular Biology of the Cell

Article Title: Reversible assembly and disassembly of V-ATPase during the lysosome regeneration cycle

doi: 10.1091/mbc.e23-08-0322

Figure Lengend Snippet: FIGURE 3: Colocalization of EGFP-tagged proteins with LAMP1-mCherry or ARL8b-mCherry in stably expressing NRK cells. (A–E) Manders’ colocalization coefficients for EGFP-tagged proteins vs LAMP1-mCherry in untreated cells, cells incubated with 30 mM sucrose for 24 h or incubated with sucrose followed by 1–8 h incubation with 0.5 mg/ml invertase. Mixed stable cells used except in (D) in which a clonal LAMP1-mCherry/EGFP-RILP cell line, 86.7 ± 1.9% (3) depleted of endogenous RILP by siRNA-mediated knockdown, was used. (F) Summary of changes in mean Manders’ colocalization coefficients shown in panels A–E. (G) Manders’ colocalization coefficients (whole cell or outer shell) for ARL8b-mCherry versus RAB7a-EGFP in untreated mixed stable ARL8b-mCherry/ RAB7a-EGFP cells, cells incubated with 30 mM sucrose for 24 h or incubated with sucrose followed by 8-h incubation with 0.5 mg/ml invertase. (H) Confocal image showing RAB7a-EGFP entering lysosome reformation tubules (arrowheads) after incubating RAB7a-EGFP cells containing sucrosomes with 0.5 mg/ml invertase for 1 h (I) Manders’ colocalization coefficients for V1G1-EGFP versus LAMP1-mCherry in the presence or absence of 80 μM dynasore or 1 μM latrunculin A in otherwise untreated LAMP1- mCherry/ V1G1-EGFP mixed stable cells, cells incubated with 30 mM sucrose for 24 h or incubated with sucrose followed by 8-h incubation with 0.5 mg/ml invertase. In panels A–E, G, and I, mean ± SEM of three experiments (minimum 10 cells per experiment) shown. *, p < 0.05; **, p < 0.01, ***, p < 0.005; ****, p < 0.001.

Article Snippet: Torin 1 was from Tocris Bioscience (Bristol, UK), dynasore monohydrate from Santa Cruz Biotechnology (Dallas, TX, USA) and bafilomycin A1 from Alfa Aesar (Heysham, UK).

Techniques: Stable Transfection, Expressing, Incubation, Knockdown

Journal: Journal of Pharmaceutics

Article Title: Biological Atomic Force Microscopy for Imaging Gold-Labeled Liposomes on Human Coronary Artery Endothelial Cells

doi: 10.1155/2013/875906

Figure Lengend Snippet: The internalization process of coupled liposomes was blocked when the cells were pretreated with 80 μ M dynasore for 15 min before 60 min of incubation with “gold-liposomes.” The gold liposome clusters did not penetrate the cell membrane, remaining just attached to the external wall (arrow in (a)). The corresponding fluorescence image (b) shows positive signaling from the “gold liposomes” that were scanned. The cells were fixed in formalin 10% and imaged at 50 μ m 2 in contact mode in liquid (DNP-S fo = 12–24 kHz, k = 0.06 N/m).

Article Snippet: Dimethyl sulfoxide anhydrous 99.9% (DMSO), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), chloroform A.C.S. reagent 99.8% with amylenes as stabilizer, 2-(N-morpholino) ethanesulfonic acid (MES), collagen from calf skin, neutralbuffered formalin 10%, and dynasore monohydrate were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Incubation, Fluorescence