1. (A) Schematic representation of the conditional Lats1 locus.
(A) Schematic representation of the conditional Lats1 locus. Upon mammary-specific Cre expression, exon 4 (colored blue) is excised. (B) Genotyping of the Lats1 allele. (C) Expression of Lats1 mRNA in WT- and Lats1-CKO PyMT tumors, analyzed by RT-qPCR. Mean ± SEM of tumors from different mice. Mouse numbers indicated at the bottom; ***P-value < (D)LATS1 and LATS2 mRNA expression levels in human breast cancer tumors (TCGA-BRCA dataset). Tumors were stratified according to histological type (n = 1095). (E) Genes were ranked according to fold change between LATS1 siRNA (siLATS1) and control siRNA in transiently transfected MCF7 cells (left) or between LATS1 overexpression and vector control in ZR75-1 cells (ZR75-1/LATS1, right) and compared with differentially expressed genes in Lats1-CKO PyMT tumors (adjP-value < 0.05, FC > 1.5). (F) YAP/TAZ target genes (Dupont gene set, Dupont et al, 2011), differentially expressed in either Lats1-or Lats2-CKO PyMT tumors relative to their WT-PyMT littermate controls (P-value < 0.05). The heat map depicts the log ratio between Lats-CKO and WT-PyMT littermate controls; grey indicates genes not significantly differentially expressed between the two conditions. (G) ZR75-1 cells were transfected with the indicated siRNA oligonucleotides. CYR61 and IGFBP3 mRNA levels were analyzed by RT-qPCR; mean ± SD of two technical replicates. (H) Functional enrichment analysis (GO biological processes) for genes differentially regulated upon LATS1 silencing in either ZR75-1 (FC > 1.5, adjP-value < 0.05, n = 733, left panel) or MCF7 (FC > 1.5, adjP-value < 0.05, n = 285, right panel). The 10 most enriched terms for each cell line are shown.
Noa Furth et al. LSA 2018;1:e
© 2018 Furth et al.