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CDK7 regulates organ size and tumor progression through Yap/Taz. ( A ) Western blot analysis of Yap and CDK7 in MDA-MB-231 cells transfected with siControl or siCDK7 and infected with lentiviruses expressing the empty vector, wild-type Yap, or the S128D mutant. ( B ) Relative mRNA levels of the indicated Hippo target genes in the indicated MDA-MB-231 cells were measured by RT-qPCR. Data are means ± SD. (*) P < 0.05; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( C ) Relative cell numbers of MDA-MB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap were measured by the WST-1 assay at different time points after infection. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( D , E ) Transwell invasion assay of MDAMB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( F ) Western blot analysis of Yap in MDA-MB-231 cells treated with <t>THZ1.</t> ( G ) Relative mRNA levels of the indicated Hippo target genes in MDA-MB-231 cells treated with THZ1. Data are means ± SD. (***) P < 0.001 (Student's t -test). ( H ) Tumor size of the indicated MDA-MB-231 xenografts in mice treated with vehicle or THZ1 at the indicated time after drug treatment. ( I ) Macroscopic images of tumors derived from the indicated MDA-MB-231 xenografts at the end of drug treatment. ( J ) Quantification of tumor weight of the indicated MDA-MB-231 xenografts at the end of drug treatment. Data are means ± SD. n = 7 mice for each group. (NS) not significant; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( K ) MST1/2 DKO mice were treated with vehicle or THZ1 (10 mg/kg) every other day starting at the age of 1 mo until 3 mo old. Representative macroscopic images of livers were shown. ( L ) Quantitative analysis of liver-to-body weight ratio. Data are means ± SD. n = 6 mice for each group. (***) P < 0.001 (Student's t -test). ( M ) Western blot analysis of control and THZ1-treated liver tissues of the indicated genotypes using the indicated antibodies. ( N , O , Q ) Liver cross-sections of the indicated genotypes treated with vehicle or THZ1 were subjected to immunostaining with anti-EPCAM, SOX9, F4/80, or Ki67 antibody. Scale bar, 100 μm. ( P ) Quantification of Ki67 + cells in liver sections of the indicated genotypes treated with vehicle or THZ1. Data are means ± SD. n = 5 sections for each group. (***) P < 0.001 (Student's t -test).
Thz1 A8882, supplied by ApexBio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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CDK7 regulates organ size and tumor progression through Yap/Taz. ( A ) Western blot analysis of Yap and CDK7 in MDA-MB-231 cells transfected with siControl or siCDK7 and infected with lentiviruses expressing the empty vector, wild-type Yap, or the S128D mutant. ( B ) Relative mRNA levels of the indicated Hippo target genes in the indicated MDA-MB-231 cells were measured by RT-qPCR. Data are means ± SD. (*) P < 0.05; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( C ) Relative cell numbers of MDA-MB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap were measured by the WST-1 assay at different time points after infection. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( D , E ) Transwell invasion assay of MDAMB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( F ) Western blot analysis of Yap in MDA-MB-231 cells treated with THZ1. ( G ) Relative mRNA levels of the indicated Hippo target genes in MDA-MB-231 cells treated with THZ1. Data are means ± SD. (***) P < 0.001 (Student's t -test). ( H ) Tumor size of the indicated MDA-MB-231 xenografts in mice treated with vehicle or THZ1 at the indicated time after drug treatment. ( I ) Macroscopic images of tumors derived from the indicated MDA-MB-231 xenografts at the end of drug treatment. ( J ) Quantification of tumor weight of the indicated MDA-MB-231 xenografts at the end of drug treatment. Data are means ± SD. n = 7 mice for each group. (NS) not significant; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( K ) MST1/2 DKO mice were treated with vehicle or THZ1 (10 mg/kg) every other day starting at the age of 1 mo until 3 mo old. Representative macroscopic images of livers were shown. ( L ) Quantitative analysis of liver-to-body weight ratio. Data are means ± SD. n = 6 mice for each group. (***) P < 0.001 (Student's t -test). ( M ) Western blot analysis of control and THZ1-treated liver tissues of the indicated genotypes using the indicated antibodies. ( N , O , Q ) Liver cross-sections of the indicated genotypes treated with vehicle or THZ1 were subjected to immunostaining with anti-EPCAM, SOX9, F4/80, or Ki67 antibody. Scale bar, 100 μm. ( P ) Quantification of Ki67 + cells in liver sections of the indicated genotypes treated with vehicle or THZ1. Data are means ± SD. n = 5 sections for each group. (***) P < 0.001 (Student's t -test).

Journal: Genes & Development

Article Title: CDK7 regulates organ size and tumor growth by safeguarding the Hippo pathway effector Yki/Yap/Taz in the nucleus

doi: 10.1101/gad.333146.119

Figure Lengend Snippet: CDK7 regulates organ size and tumor progression through Yap/Taz. ( A ) Western blot analysis of Yap and CDK7 in MDA-MB-231 cells transfected with siControl or siCDK7 and infected with lentiviruses expressing the empty vector, wild-type Yap, or the S128D mutant. ( B ) Relative mRNA levels of the indicated Hippo target genes in the indicated MDA-MB-231 cells were measured by RT-qPCR. Data are means ± SD. (*) P < 0.05; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( C ) Relative cell numbers of MDA-MB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap were measured by the WST-1 assay at different time points after infection. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( D , E ) Transwell invasion assay of MDAMB-231 cells treated with siControl or siCDK7 and infected with lentiviruses expressing wild-type or S128D Yap. Data are means ± SD. (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( F ) Western blot analysis of Yap in MDA-MB-231 cells treated with THZ1. ( G ) Relative mRNA levels of the indicated Hippo target genes in MDA-MB-231 cells treated with THZ1. Data are means ± SD. (***) P < 0.001 (Student's t -test). ( H ) Tumor size of the indicated MDA-MB-231 xenografts in mice treated with vehicle or THZ1 at the indicated time after drug treatment. ( I ) Macroscopic images of tumors derived from the indicated MDA-MB-231 xenografts at the end of drug treatment. ( J ) Quantification of tumor weight of the indicated MDA-MB-231 xenografts at the end of drug treatment. Data are means ± SD. n = 7 mice for each group. (NS) not significant; (**) P < 0.01; (***) P < 0.001 (Student's t -test). ( K ) MST1/2 DKO mice were treated with vehicle or THZ1 (10 mg/kg) every other day starting at the age of 1 mo until 3 mo old. Representative macroscopic images of livers were shown. ( L ) Quantitative analysis of liver-to-body weight ratio. Data are means ± SD. n = 6 mice for each group. (***) P < 0.001 (Student's t -test). ( M ) Western blot analysis of control and THZ1-treated liver tissues of the indicated genotypes using the indicated antibodies. ( N , O , Q ) Liver cross-sections of the indicated genotypes treated with vehicle or THZ1 were subjected to immunostaining with anti-EPCAM, SOX9, F4/80, or Ki67 antibody. Scale bar, 100 μm. ( P ) Quantification of Ki67 + cells in liver sections of the indicated genotypes treated with vehicle or THZ1. Data are means ± SD. n = 5 sections for each group. (***) P < 0.001 (Student's t -test).

Article Snippet: Mice were treated with THZ1 (APEXBIO, catalog no. A8882) by intraperitoneal injection at a concentration of 10 mg/kg every other day or in equivalent volumes of vehicle as a control.

Techniques: Western Blot, Transfection, Infection, Expressing, Plasmid Preparation, Mutagenesis, Quantitative RT-PCR, WST-1 Assay, Transwell Invasion Assay, Derivative Assay, Control, Immunostaining

Targeting CDK7 for treating Yap/Taz-driven cancer. ( A ) A conserved role of CDK7/CRL4 DCAF12 in the regulation of nuclear Yki/Yap/Taz. CDK7 phosphorylates Yki/Yap/Taz to prevent its binding to CRL4 DCAF12 , thereby stabilizing Yki/Yap/Taz in the nucleus. ( B , top ) Yap/Taz/TEAD binds superenhancers together with other transcriptional factors (X, Y, etc.) to drive the expression of genes essential for tumor growth. ( Bottom ) CDK7 inhibitor THZ1 blocks tumor growth by inhibiting both Yap/Taz and Pol II in Yap/Taz-driven cancer.

Journal: Genes & Development

Article Title: CDK7 regulates organ size and tumor growth by safeguarding the Hippo pathway effector Yki/Yap/Taz in the nucleus

doi: 10.1101/gad.333146.119

Figure Lengend Snippet: Targeting CDK7 for treating Yap/Taz-driven cancer. ( A ) A conserved role of CDK7/CRL4 DCAF12 in the regulation of nuclear Yki/Yap/Taz. CDK7 phosphorylates Yki/Yap/Taz to prevent its binding to CRL4 DCAF12 , thereby stabilizing Yki/Yap/Taz in the nucleus. ( B , top ) Yap/Taz/TEAD binds superenhancers together with other transcriptional factors (X, Y, etc.) to drive the expression of genes essential for tumor growth. ( Bottom ) CDK7 inhibitor THZ1 blocks tumor growth by inhibiting both Yap/Taz and Pol II in Yap/Taz-driven cancer.

Article Snippet: Mice were treated with THZ1 (APEXBIO, catalog no. A8882) by intraperitoneal injection at a concentration of 10 mg/kg every other day or in equivalent volumes of vehicle as a control.

Techniques: Binding Assay, Expressing