Extended Data Fig. 3: MCB and VT01454 preferentially kills YAP dependent cancer cells.
From: Hippo pathway regulation by phosphatidylinositol transfer protein and phosphoinositides
a and b, MCB induces cell death in the GNAQ/11 mutant 92.1 and OMM1, but not the B-Raf mutant OCM1 and OCM8 cells where VP displays general toxicity. The concentrations of MCB and VP were 2.7μM and 2 μM, respectively. Scale bar = 50 μm. Results are mean ± SD of three biologically independent samples. c, 92.1 cells are more sensitive than OCM1 cells to MCB-induced apoptosis. The MCB concentrations were 2.7 and 5.4 μM. d, MCB is more selective than verteporfin to induce PARP cleavage. Concentration gradients (0, 0.27, 0.68, 1.36, 2.72, 6.8 μM) of MCB or verteporfin (0, 0.28, 0.70, 1.39, 2.78, 6.9 μM) were used to treat cells. PARP cleavage was determined by Western blot. e, MCB (2.7 μM) preferentially inhibits colony formation in 92.1 cells over OCM1 cells. Both images and quantification of colonies in soft agar assays are shown. Results are mean ± SD of three biologically independent samples. f, VT01454 does not induce YAP-5SA phosphorylation. The stable 92.1 cell pools were generated by infection with lentivirus expressing YAP-5SA or empty-vector. These cells were treated with 20 nM or 50 nM of VT01454 for 1 hours before subjecting to western blot with indicated antibodies. g, VT01454 sensitivity in multiple cancer cell lines. 1.5 × 105 cells were seeded and treated with VT01454 (20 nM) or DMSO for 48 hours. Scale bar = 100 μm. Representative images were taken by a phase-contrast microscope. h, Quantification of live cell numbers (left panel) was performed by cell counting after trypan blue staining. The right panel was the normalized result of the left panel by setting the DMSO control as 1. Experiments were similar to panel g. Results were mean ± SD of three biologically independent samples.
