Fig. 6

Validation of iBET151 and iBET762 on MDCK p60AmotL2-expressing cells in 3D gels. A and B Dose‒response viability testing of MDCK cells grown in 3D Matrigel with CTG after 72 h of treatment with iBET151 (A) and iBET762 (B), respectively. The IC50 values were calculated using the dose‒response nonlinear regression model from GraphPad Prism. The percentage of viability for p60AmotL2-expressing cells is represented by the purple line, and the values are compared to controls. From top to bottom, 95% CI for iBET151 (A): [5.992, 14.82], [4.748, 8.732], [2.950, 7.570] and [0.3647, 0.6111]; 95% CI for iBET762 (B): [6.073, 11.23], [5.154, 8.674], [4.626, 8.594] and [1.611, 2.748]. C IF confocal imaging of MDCK cells grown in 3D collagen. Cells were cultured in a collagen matrix for approximately 7 days, treated with Dox for 48 h to induce p60AmotL2 expression, and exposed to HGF for 24 h to induce migration of control cells and scattering of p60AmotL2-expressing cells. Subsequently, cells were treated with iBET151 for 24 to 48 h before fixation. Cleaved caspase-3 staining (in green) was used to assess cell inhibition or cell death; white arrows indicate migrating cells positive for cleaved caspase-3 following iBET151 treatment. Data represent the mean ± SEM from three independent experiments. Scale bars = 50 μm