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. 2018 Dec 7;8:17705. doi: 10.1038/s41598-018-35792-8

Figure 6.

Figure 6

CASP4-silencing impairs cell detachment and influences number and size of focal adhesions. (a,b) Percentage of cells detached from tissue culture plate upon treatment with trypsin/EDTA is shown in y-axis. The percentage is calculated with respect to untreated LR1.2, LR3.2 and LR4.2 cell lines (a) and untreated siCTRL and siCASP4 transfected A431 cells (b). Time (min) of incubation in detachment solution is reported in x-axis. Data are representative of three independent experiments (LR1.2 - LR3.2 and LR1.2 - LR4.2 at 2, 4, 10 and 15 min, p = 0.05, n = 3–6; siCASP4 - siCTRL: 4 min, p = 0.04; 10 min, p = 0.05, n = 3). (c) Representative confocal microscopy images of talin-stained (red) cells, cultured on poly-L-lysine treated glasses; 4x enlargments are also shown, corresponding to the yellow boxes in LR1.2 (A), LR3.2 (B) and LR4.2 (C). The bar plots indicate the percentage of focal adhesions (FA) positive cells (n = 34–38), the number of FA per cell (n = 39–42) and the FA length (n = 236–597) calculated in clusters of 6–10 cells present in 7–10 fields from three independent experiments (FA positive cells, FA number/cells, FA length: LR1.2 - LR3.2 and LR1.2 - LR4.2, p < 0.0001; FA number/cells: LR3.2 - LR4.2, p = 0.03). Scale bars are indicated. Statistical analysis for every pair-comparison was performed by Wilcoxon rank sum test. Significant p-values are represented by asterisks: *p < 0.05; ****p < 0.0001. Non-significant p-values are not shown.