Journal: American Journal of Cancer Research

Article Title: Modifications of disulfide bonds in breast cancer cell migration and invasiveness

doi:

Figure Lengend Snippet: Adhesion of MCF-10A (A), MCF-7 (B) and MDA-MB-231 (C) Cells to collagen type I and vitronectin in the presence of PDI inhibitors and free thiol blockers. Data presented as mean + SD; mini-max ranges marked as whiskers; n=6. Total numbers of adherent cells were estimated with BCA Protein Assay. Significance of differences was analysed with one-way ANOVA and the post-hoc multiple comparisons Tukey’s test and planned comparisons were verified the bootstrap-boosted unpaired student’s t test (10000 iterations) with the Bonferroni’s correction for multiple comparisons.

Article Snippet: Materials Horseradish conjugated secondary antibodies were purchased from Santa Cruz biotechnology (Santa Cruz, CA, USA), Alexa Fluor conjugated antibodies, antibodies anti-β 1 clone 12G10 were purchased from Abcam (Cambridge, UK), chemiluminescence detection system was purchased from Cyanagen (Bologna, BO, Italy), antibodies anti-PDI RL77, Pierce BCA Protein Assay Kit, Pierce IP Lysis Buffer, Pierce Control Agarose, protein A/G-agarose, CellTracker Green CMFDA Dye, protease inhibitor cocktail and neutravidin-agarose were purchased from Thermo Scientific Pierce (Waltham, MA, USA), GSH, iodoacetamide, DTNB, MPB, cystamine, rutin hydrate (Q3Rut), 16F16, PACMA-31, NEM, TCEP were purchased from Sigma-Aldrich LTD (St. Louis, MO), vitronectin, antibodies anti-β 1 AB1952 and anti-β 1 MAB1987, anti-α V β 3 clone LM609 and anti-α V β 3 clone CBL544 were purchased from Merck Milipore (Billerica, MA, USA); Bovine Collagen Solution PureCol was purchased from Advanced BioMatrix Inc. (San Diego, CA, USA); all standard tissue culture reagents, including DMEM, fetal bovine serum, trypsin/EDTA and others were purchased from GIBCO (Waltham, MA, USA).

Techniques: Bicinchoninic Acid Protein Assay

Journal: American Journal of Cancer Research

Article Title: Modifications of disulfide bonds in breast cancer cell migration and invasiveness

doi:

Figure Lengend Snippet: Migration of MCF-10A (A), MCF-7 (B) and MDA-MB-231 (C) Cells to collagen type I and vitronectin in the presence of PDI inhibitors and free thiol blockers. Data presented as mean + SD; mini-max ranges marked as whiskers; n=6. Pictures of overgrowing scratches were taken following 24-hour incubation with the tested inhibitors/blockers using an inverted light microscopy. Significance of differences was analysed with one-way ANOVA and the post-hoc multiple comparisons Tukey’s test and planned comparisons were verified the bootstrap-boosted unpaired student’s t test (10000 iterations) with the Bonferroni’s correction for multiple comparisons.

Article Snippet: Materials Horseradish conjugated secondary antibodies were purchased from Santa Cruz biotechnology (Santa Cruz, CA, USA), Alexa Fluor conjugated antibodies, antibodies anti-β 1 clone 12G10 were purchased from Abcam (Cambridge, UK), chemiluminescence detection system was purchased from Cyanagen (Bologna, BO, Italy), antibodies anti-PDI RL77, Pierce BCA Protein Assay Kit, Pierce IP Lysis Buffer, Pierce Control Agarose, protein A/G-agarose, CellTracker Green CMFDA Dye, protease inhibitor cocktail and neutravidin-agarose were purchased from Thermo Scientific Pierce (Waltham, MA, USA), GSH, iodoacetamide, DTNB, MPB, cystamine, rutin hydrate (Q3Rut), 16F16, PACMA-31, NEM, TCEP were purchased from Sigma-Aldrich LTD (St. Louis, MO), vitronectin, antibodies anti-β 1 AB1952 and anti-β 1 MAB1987, anti-α V β 3 clone LM609 and anti-α V β 3 clone CBL544 were purchased from Merck Milipore (Billerica, MA, USA); Bovine Collagen Solution PureCol was purchased from Advanced BioMatrix Inc. (San Diego, CA, USA); all standard tissue culture reagents, including DMEM, fetal bovine serum, trypsin/EDTA and others were purchased from GIBCO (Waltham, MA, USA).

Techniques: Migration, Incubation, Light Microscopy