Fig. 8

Synthetic lethality regulated by Bcl-xL and YAP signaling. A Sphere-forming assay using control, Rasa1-KO, and Rasa1/Nf2-double-KO S1 cells treated with BH3I-1 (25 µM) for 5 days. B and C Effect of Bcl-xL inhibition on peritoneal dissemination model of Rasa1/Nf2-double-KO S1M cells in NOD-SCID mice treated with vehicle (n = 5) or A-1155463 (n = 5, 7.5 mg/kg, i.p., once daily) for 10 consecutive days. B Ascites volume and (C) macro-metastatic foci numbers measured at necropsy. D HOP-Flash luciferase reporter assay in BH3I-1-treated S1 cells after 48 h. E, F, and G Sphere-forming assay of Rasa1/Nf2-double-KO S1 cells treated with BH3I-1 (50 µM) and/or verteporfin (1 µM) for 5 days. E Tumorsphere size, (F) number, and (G) Representative images of tumorspheres. H Luciferase ATP cell viability assay of Rasa1/Nf2-double-KO S1 tumorspheres, treated with BH3-I and/or verteporfin for 5 days at indicated doses. I RT-qPCR analysis of mRNA expression levels of (top) Wnt-dependent transcription (Ccnd1) and (bottom) YAP-dependent transcription (Cyr61) depending on BH3-I and/or verteporfin treatment in Rasa1/Nf2-double-KO S1 tumorspheres. J-M (J) Bioluminescence in vivo imaging of NOD-SCID mice 8 days after intraperitoneal transplantation of in Rasa1/Nf2-double-KO S1M cells, treated with vehicle (n = 5), A-1155463 (n = 5, 7.5 mg/kg, i.p., once daily), verteporfin (n = 5, 10 mg/kg, i.p., every other day), or combination of both drugs (n = 5). K Total bioluminescence signals in (J). L Representative H&E-stained images of peritoneal metastatic foci. Bar = 200 μm. M Depth of invasion was measured based on H&E slides. N, O, and P (N) Subcutaneous tumor volume of Rasa1/Nf2-double-KO S1M cells treated with vehicle (n = 5), A-1155463 (n = 5, 7.5 mg/kg, i.p., once daily), verteporfin (n = 5, 10 mg/kg, i.p., every other day) or combination of both drugs (n = 5). O Representative histopathological images of pulmonary metastasis. Black arrows indicate metastatic foci. Red boxes represent magnified views of red arrow locations. Bar = 1 mm (P) Statistical analysis of pulmonary metastasis regarding the number of metastatic foci (top) and mean foci size (bottom). Student's t-test was used for statistical analysis, unless otherwise specified