Extended Data Fig. 1: LATS1/2 are required to maintain ERα and target-gene expression.
From: Hippo signalling maintains ER expression and ER+ breast cancer growth
a, Impaired YAP phosphorylation in LATS1/2 double-knockout cells. Wild-type MCF-7 cells, and two MCF-7 LATS1/2 double-knockout clones, were serum-starved or treated with 1 μM cerivastatin (for 1 h), and subjected to immunoblot analysis. b, Increased YAP and TAZ transcriptional activity in LATS1/2 double-knockout cells. qPCR of the YAP and TAZ target-genes CTGF and CYR61 (also known as CCN1). c, LATS1/2 deficiency inhibits growth of MCF-7 cells. d, YAP signature and oestrogen receptor signature are among the top upregulated and downregulated gene sets, respectively, in LATS1/2 double-knockout cells. Gene enrichment analysis of LATS1/2-deficient (n = 3) and wild-type (n = 3) MCF-7 cells. Circle size represents the relative gene numbers in each set. Red, enriched in LATS1/2-deficient cells; blue, enriched in wild-type cells. e, Opposite effects of LATS1/2 deletion on the YAP signature genes (red) and oestrogen response signature genes (blue) in MCF-7 cells. Significance, false-discovery rate-adjusted P value. Magnitude of difference, fold change. n = 3 independent samples. f, YAP, TAZ and ERα localization and intensity in wild-type or LATS1/2 double-knockout MCF-7 cells. Scale bar, 20 μm. g, h, Redundancy of LATS1/2 in regulating YAP activity and ERα expression. MCF-7 clones with different sgRNA or shRNA targeting LATS1, LATS2 or both were subjected to qPCR analysis for CTGF and ESR1 (g) or immunoblot (h). i–k, Deletion of Lats1/2 reduces ERα in mouse endometrial organoids. Organoids derived from the endometrial tissues of Lats1+/+Lats2+/+ and Lats1fl/flLats2fl/fl mice were infected with Cre-encoding adenovirus and subjected to immunohistochemistry (i), immunoblot (j) and qPCR analysis (k). Scale bar, 100 μm for bright field, 30 μm for immunohistochemistry. l–n, Deletion of Lats1/2 reduces ERα in mouse Fallopian tube organoids. Organoids derived from the Fallopian tube tissues of Lats1+/+Lats2+/+ and Lats1fl/f/Lats2fl/fl mice were infected with Cre-encoding adenovirus and subjected to immunohistochemistry (l), immunoblot (m) and qPCR analysis (n). Scale bar, 200 μm for bright field, 25 μm for immunohistochemistry staining. ***P < 0.001; mean + s.d.. Gel source data are provided in Supplementary Fig 1.
