Journal: Oncotarget

Article Title: Eribulin targets a ch-TOG-dependent directed migration of cancer cells

doi:

Figure Lengend Snippet: Following treatment with eribulin for 4 h, SKBr3 cells were stimulated with HRG for 30 min and fixed. EB1, CLIP-170 and tubulin localizations were visualized by triple immunofluorescence labeling. White scale bars represent 10 μm; cell limits are delineated with dashed lines.

Article Snippet: In the case of ch-TOG labeling, cells were simply fixed with methanol at −80°C for 10 min. Immunolabeling was performed with antibodies against EB1 (Santa Cruz Biotechnology, Dallas, TX), CLIP-170 (Synaptic Systems, Goettingen, Germany), α-tubulin (Sigma-Aldrich) or ch-TOG (a generous gift from Dr Lynn Cassimeris, Lehigh University Bethlehem, PA) and secondary antibodies labeled with DyeLight ® 405, AlexaFluor ® 488 or 594 (Jackson ImmunoResearch Europe, Suffolk, UK).

Techniques: Immunofluorescence, Labeling

Journal: Oncotarget

Article Title: Eribulin targets a ch-TOG-dependent directed migration of cancer cells

doi:

Figure Lengend Snippet: The length and density of fluorescent EB1 and corresponding CLIP-170 comets were measured on immunofluorescence images such as depicted in Figure . A. For each condition, a total of 90 comets from three independent experiments (10 comets per cell, 3 cells per experiment) were measured. B. For each condition, a total of 13 cells from five independent experiments were analyzed for the number of EB1 or CLIP-170 comets in an area of 300 μm 2 . C. EB1-GFP comets were tracked in HRG-stimulated SKBr3 cells, pre-treated or not with 0.1 nM eribulin for 4 h, and resulting tracks were analyzed using +TipTracker. Growth speed (including pauses) and growth lifetime were calculated. For each condition, a total of >14,000 tracks, in 15 cells from 3 independent experiments were analyzed. Medians and SD are presented, Student t-test with Welch correction: ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In the case of ch-TOG labeling, cells were simply fixed with methanol at −80°C for 10 min. Immunolabeling was performed with antibodies against EB1 (Santa Cruz Biotechnology, Dallas, TX), CLIP-170 (Synaptic Systems, Goettingen, Germany), α-tubulin (Sigma-Aldrich) or ch-TOG (a generous gift from Dr Lynn Cassimeris, Lehigh University Bethlehem, PA) and secondary antibodies labeled with DyeLight ® 405, AlexaFluor ® 488 or 594 (Jackson ImmunoResearch Europe, Suffolk, UK).

Techniques: Immunofluorescence