Fig. 5

HTH-02-006 attenuates YAP-driven cell proliferation, hepatomegaly and tumorigenesis. a Chemical structures and IC50 of HTH-02-006. b In the presence of 100 μM [γ-32P]ATP, NUAK2 activity was analyzed using 200 μM Sakamototide with the indicated concentrations of HTH-02-006. The results are presented as the percentage of kinase activity relative to the DMSO-treated control. Results are mean ± S.D. n = 2. c Western blot analysis of phospho-S445 MYPT1, MYPT1, phospho-MLC, and MLC in SNU475 cells treated with HTH-02-006. d Cell growth curves of indicated liver cancer cell lines. Cells were treated with increasing concentrations of HTH-02-006 for 120 h. Data are presented as mean ± SD; n = 3. e Western blot analysis of indicated antibodies in wild-type and A236T mutant cells treated with/without HTH-02-006. f Growth curves of HuCCT-1 wild-type and A236T mutant cells treated with increasing concentrations of HTH-02-006 for 120 h. Data are presented as mean ± SD; n = 3. g TetO-YAP S127A mice infected with AAV-Cre were fed Dox for 14 days. Data are presented as mean ± SD; n = 7 animals per group. HTH-02-006 was administered by intraperitoneal injection twice a day during the 14 days period. Gross morphology of the liver in TetO-YAP S127A mice treated with HTH-02-006. Liver/body weight ratio of TetO-YAP S127A mice at the end of period is shown. Ki67 staining of liver sections and quantification of Ki67 positive cells are shown. Data are presented as mean ± SD; n = 3, 5 HPF per mouse. The two-tailed, Student’s t test was used to compare between two groups and expressed as P values. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. h Immunoblotting of liver lysates from TetO-YAP S127A mice treated for 14 days with vehicle or HTH-02-006. Three independent samples from each treatment were used. Each lane represents a different mouse liver. i Tumor growth of HuCCT-1 TetO-YAP S127A cells in a xenograft model. Nude mice were fed with or without Dox. HTH-02-006 (10 mg/kg) was administered by intraperitoneal injection twice a day. Data are presented as mean ± SD; n = 4 animals per group