Fig. 5

HCAb2 reduced in vitro migration of MDA-MB-231 cells. a-f Representative images of scratch assay, T = 0 h (panels a-c) and T = 19 h (panels d-f). Scratches were made using 200 μL pipet tips and T = 0 h images were taken. Subsequently cells were left untreated or incubated with HCAb1 and HCAb2 (5 μg each) and imaged after 19 h. Scale bar represents 100 μm. g Quantification of percent acellular area remaining after 19 h of treatment with HCAb1 and HCAb2. Acellular area at T = 0 h and T = 19 h was determined for each well using Image-Pro software (n = 4 wells per treatment). Average area at T = 0 h for each treatment was set to be 100 % and areas at T = 19 h were normalized to the corresponding average acellular area at T = 0 h. Percent acellular area remaining was calculated accordingly. Error bars represent standard deviation and statistical significance was determined by Student’s t test, * = p ≤ 0.05. h Quantification of cell migration in a transwell assay. MDA-MB-231 cells were left untreated or pre-treated with 10 μg of HCAb1 or HCAb2 or anti-HSP90 antibody (sc-1055) or 10 μM 17-DMAG for 15 min at room temperature. Cells were allowed to migrate for 19 h at 37 °C with EGF as the chemoattractant. Cells were stained with crystal violet and total numbers of migrated cells were counted. Average number of migrated cells with standard deviation was plotted for 3 transwell inserts per treatment. Statistical significance was determined using Student’s t test, ** = p ≤ 0.01