MDA‐MB‐231 cells stably expressing a lentiviral hairpin targeting USP26 (L1) or relevant control vector were plated for scratch assay and treated with either SB431542 (5 μM) or TGF‐β (5 ng/ml); panels show migration at 0 and 24 h. Representative images are shown (scale bars, 50 μm).
Percentage of migrated area within the white dotted lines was estimated with respect to control (0 h) and a graph was plotted. ***P < 0.001 using Student's t‐test. Data are mean ± SD of three random fields. Data are representative of two independent experiments with similar results.
Transwell assay of MDA‐MB‐231 cells stably expressing a lentiviral hairpin targeting USP26 (L1) or relevant control vector treated with SB431542 (5 μM) or TGF‐β (5 ng/ml) for 16 h prior to fixation and crystal violet staining. Representative images are shown (scale bars, 100 μm).
Graph represents average number of migrated cells taken from four different random fields from panel (C). Data are mean ± SD of triplicate samples from a representative experiment performed three times. ***P < 0.001 using Student's t‐test.
Transwell assay of MDA‐MB‐231 cells stably expressing GFP‐USP26, GFP‐USP26 C/S, or relevant control vector treated with SB431542 (5 μM) or TGF‐β (5 ng/ml) for 16 h prior to fixation and crystal violet staining. Representative images are shown (scale bars, 100 μm).
Graph represents average number of migrated cells taken from four different random fields from panel (E). Data are mean ± SD of triplicate samples from a representative experiment performed three times. ***P < 0.001 using Student's t‐test.
Immunoblotting showing ectopically expressed GFP‐USP26, GFP‐USP26 C/S, or control vector in MDA‐MB‐231.