Fig. 2

Effect of LOXL1 expression on the migration and invasion of HCT8 and SW480 cells in vitro. a Western blot analysis demonstrating the expression of LOXL1 in CRC cell lines. Total GAPDH was used as a loading control. b Western blot analysis of HCT8/SW480 cells stably transfected with LOXL1 overexpression lentiviruses or control lentiviruses. Total GAPDH served as a loading control. c Wound healing analysis: effect of the overexpression of LOXL1 in stable HCT8 cells at 0 h, 24 h, and 48 h (upper panel) and calculation of the percentage of wound healing (lower panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. ***P < 0.001. Scale bar: 100 μm. d Transwell migration and Matrigel invasion assays using stable HCT8-LOXL1 and HCT8-N cells (left panel) and calculation of the rate of migration/invasion in relevant stable HCT8 cells (right panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. *P < 0.05, ***P < 0.001. Scale bar: 100 μm. e A colony formation assay was performed in HCT8 cells with or without LOXL1 overexpression. Left panel: representative images, right panel: quantification analysis. Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. ***P < 0.001. f Wound healing analysis to determine the effect of LOXL1 overexpression in stable SW480 cells at 0 h, 24 h, and 48 h (upper panel) and calculation of their wound healing percentages (lower panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. *P < 0.05, ****P < 0.0001. Scale bar: 100 μm. g Transwell migration and Matrigel invasion assays using stable SW480-LOXL1 and SW480-N cell lines (left panel) and calculation of the rate of migration/invasion in corresponding stable SW480 cells (right panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. ***P < 0.001. Scale bar: 100 μm. h A colony formation assay was performed using SW480 cells with or without LOXL1 overexpression. Left panel: representative images, right panel: quantification analysis. Data from independent experiments are presented as the mean ± SD. Statistical significance was assessed by unpaired t-test; ***P < 0.001.