LATS2 promotes death of lumB cells.

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LATS2 promotes death of lumB cells. LATS2 promotes death of lumB cells. (A) ZR75-1 cells were treated with 1 μM 5-aza-2′-deoxycytidine (5-Aza) for 4 d. Upper panel: RT-qPCR analysis of LATS2 mRNA; mean ± SD of two technical replicates. Lower panel: Western blot analysis of LATS2 protein. (B) Functional enrichment of cell viability-related terms for differentially expressed genes in luminal cancer cell lines with transient silencing (siLATS2) or stable overexpression (OE) of LATS2, compared with controls. Ingenuity Pathway Analysis (IPA); Diseases or Functions annotations. (C) ZR75-1 cells were transiently transfected with GFP-LATS1 or GFP-LATS2, stained with anti-cleaved caspase 3 (CC3) antibody after 48 h, and subjected to imaging flow cytometry (ImageStreamX). Only cells with intact nucleus and positive GFP signal were analyzed; mean ± SEM of staining intensity; ***P-value < 0.001. (D) ZR75-1 cells were transfected with GFP-LATS1 or GFP-LATS2 or GFP only (vector). 72 h later, cell viability was assessed by PI exclusion followed by flow cytometry analysis. “Cell death” represents the ratio between the percentages of dead cells (PI+) in the GFP-positive population versus the GFP-negative population; mean ± SEM of three independent experiments; ***P-value < 0.001. Representative FACS results are shown on the right; in each case, upper and lower panels represent the GFP-positive and GFP-negative subpopulation, respectively, of the same transfected culture. (E) ZR75-1 cells stably transduced with MYC-LATS2 plasmid or control vector were subjected to PI exclusion analysis followed by flow cytometry. Left panel: percentage of dead (PI+) cells. Right panel: cell count of live (PI−) cells; mean ± SEM of three independent experiments; *P-value < 0.05, **P-value < 0.01. (F) Histological samples from carcinomas of Lats2-CKO PyMT and WT-PyMT littermate controls were immunostained for γH2AX and cleaved caspase 3 (CC3, left panel, scale bar = 100 μm). Right panel: mean ± SEM of percentage of positive cells, based on at least nine sections of each genotype; ***P-value < 0.001. Noa Furth et al. LSA 2018;1:e201800171 © 2018 Furth et al.