Journal: Oncotarget

Article Title: Inhibitors of Rho kinase (ROCK) signaling revert the malignant phenotype of breast cancer cells in 3D context

doi: 10.18632/oncotarget.9395

Figure Lengend Snippet: A. Morphological alternations of T4-2 cells treated with ROCK inhibitor (Y-26732 or Fasudil, 30 μM), EGFR inhibitor (AG1478, 0.1 μM), Integrinβ1 function blocking antibody (AIIB, 100 μg/mL), EGFR function blocking antibody (mAB225, 4 μg/mL) or MEK inhibitor (PD98059, 10 μM). Scale bars: 20 μm. B. C. and D. Confocal immunofluorescence of basal maker, lateral marker and cytoskeletal filamentous F. -actin. Their images were obtained using their specific antibodies ( B. Integrinα6; Green, C. β-catenin; Green) and Alexa Fluor-conjugated phalloidin ( D. ) F-actin; Red). Nuclei were stained with DAP (Blue). Scale bars: 10 μm.

Article Snippet: After blocking with immunofluorescence buffer (0.2% Triton X-100, 0.05% Tween 20, 0.1% BSA, 7.7 mM NaN 3 in PBS) containing 10% goat serum and 1% goat F(ab')2 anti-mouse immunoglobulin G (IgG; Caltag) at room temperature for 30 min, colonies were stained with anti-Integrinα6 antibody (BD phamingen) or anti-β-catenin antibody (BD phamingen) at 4°C overnight.

Techniques: Blocking Assay, Immunofluorescence, Marker, Staining

Journal: Oncotarget

Article Title: Inhibitors of Rho kinase (ROCK) signaling revert the malignant phenotype of breast cancer cells in 3D context

doi: 10.18632/oncotarget.9395

Figure Lengend Snippet: A. Morphology of MCF10A transfected with vector or pCAG-myc-ROCKΔ3 expressing constructively active ROCK. Percentage of disorganized colonies per total colonies was calculated using the amount of 4 independent images in each well. Values represent means ± SE of five determinations. *** P < 0.001 compared with vector control group (Student's t). B. Representative confocal images of DAPI-stained vector and ROCKΔ3 expressed MCF10A colonies. An arrow shows the direction from edge (left) to center (right) of colony. White scale bars: 10 μm. C. Confocal immunofluorescence of basal maker were obtained using anti-Integrinα6 antibody (Green).

Article Snippet: After blocking with immunofluorescence buffer (0.2% Triton X-100, 0.05% Tween 20, 0.1% BSA, 7.7 mM NaN 3 in PBS) containing 10% goat serum and 1% goat F(ab')2 anti-mouse immunoglobulin G (IgG; Caltag) at room temperature for 30 min, colonies were stained with anti-Integrinα6 antibody (BD phamingen) or anti-β-catenin antibody (BD phamingen) at 4°C overnight.

Techniques: Transfection, Plasmid Preparation, Expressing, Staining, Immunofluorescence

Journal: Oncotarget

Article Title: Inhibitors of Rho kinase (ROCK) signaling revert the malignant phenotype of breast cancer cells in 3D context

doi: 10.18632/oncotarget.9395

Figure Lengend Snippet: A. Morphological alternations of MDA-MB-231 cells treated with ROCK inhibitor (Y-26732 or Fasudil, 30 μM). Scale bars: 20 μm. B. Partial inhibition of proliferation of MDA-MB-231 cells treated with ROCK inhibitor (Y-26732 or Fasudil, 30 μM), EGFR inhibitor (AG1478, 0.1 μM) and MEK inhibitor (PD98059, 10 μM). Cell viability was assessed by MTT assay. Values represent means ± SE of four experiments. * P < 0.05, *** P < 0.001 compared with vehicle control group (Student's t). C. Protein expression of ROCK1, ROCK2, EGFR, Integrinβ1, GLUT3, RhoA, P-MLC and Lamin A/C in MDA-MB-231 cells. Total cell lysates were analyzed by Western blotting with their specific antibodies. D. Confocal immunofluorescence images of MDA-MB-231 cells treated with ROCK inhibitor (Y-26732 or Fasudil, 30 μM). Their images were obtained using specific antibody or Alexa Fluor-conjugated phalloidin (Integrinα6; Green, F-actin; Red, Nuclei; Blue). Scale bars: 10 μm.

Article Snippet: After blocking with immunofluorescence buffer (0.2% Triton X-100, 0.05% Tween 20, 0.1% BSA, 7.7 mM NaN 3 in PBS) containing 10% goat serum and 1% goat F(ab')2 anti-mouse immunoglobulin G (IgG; Caltag) at room temperature for 30 min, colonies were stained with anti-Integrinα6 antibody (BD phamingen) or anti-β-catenin antibody (BD phamingen) at 4°C overnight.

Techniques: Inhibition, MTT Assay, Expressing, Western Blot, Immunofluorescence