Fig. 5

Synthetic lethality regulated by Bcl-2 and YAP signaling. A Relative cell viability of control and Nf2-KO S1 tumorspheres using luciferase ATP cell viability assay. Verteporfin was treated for 5 days at indicated doses. B Sphere-forming assay using Nf2-KO and Nf2/Yap1-double-KO S1 cells. Tumorspheres diameter (left) and counts (right). C TOP-Flash luciferase reporter assay in control and Yap1-KO S1M cells. D Relative mRNA expression of Wnt-dependent transcription (Axin2) in control and Yap1-KO S1M cells. E TOP-Flash luciferase reporter assay in S1M cells, verteporfin treated for 48 h at indicated doses. F Relative mRNA expression of Wnt-dependent transcription (Aqp5 and Axin2) in S1M cells treated with verteporfin (0.5 µM) for 48 h. G Relative Bcl-2 mRNA expression in Nf2-KO S1 cells treated with verteporfin for 48 h. H TOP-Flash luciferase reporter assay in control and Yap1-KO S1 cells treated with venetoclax and/or BH3I-1 for 48 h. I, J and K Sphere-forming assay of S1 cells treated with venetoclax (5 µM) and/or verteporfin (0.4 µM) for 5 days. Tumorsphere size (I) and number (J). K Representative images of S1 tumorspheres. Bar = 200 µm. L and M Sphere-forming assay using MKN-74 cells. Venetoclax (5 µM) and/or verteporfin (0.4 µM) treated for 5 days. Tumorsphere size (L) and number (M). N Relative mRNA expression of (top) Ccnd1 and (bottom) Cyr61 in S1M cells treated with venontoclax and/or verteporfin at indicated dose for 48 h. O, P and Q Nf2-KO S1M cells were peritoneally injected in NOD-SCID mice with treatment of control (n = 5), venetoclax (n = 4, 12 mg/kg, p.o.), verteporfin (n = 5, 10 mg/kg, i.p.), and combination of venetoclax and verteporfin (n = 4). O epresentative bioluminescence imaging of NOD-SCID mice 8 days after intraperitoneal transplantation of Nf2-KO S1M cells with drug treatment. P H&E images of peritoneal metastatic tissues. Bar = 200 µm. Q Proportion of metastatic foci area (top) and depth of invasion (bottom) depending on treatment. Student's t-test was used for statistical analysis