Extended Data Fig. 1: TRIM37 overexpression in HCT116 and RPE-1 cells recapitulates synthetic lethality with centrosome loss.
From: Targeting TRIM37-driven centrosome dysfunction in 17q23-amplified breast cancer
Related to Fig. 1. a, Top, centrosome number distribution in interphase MCF-7 cells at various times after addition of centrinone (PLK4i) (125 nM). Mean ± s.e.m. Bottom, representative images of centrosome staining (centrioles labelled by centrin, and PCM labelled by CEP192). n = 3 biological replicates, each comprising >100 cells. b, Representative data of a 14-day clonogenic survival assay of MCF-7 and RPE-1 cells with the indicated genotypes treated with DMSO (control) or centrinone (PLK4i) (125 nM). n = 3 biological replicates. c, Immunoblot showing TRIM37 protein levels in two WT MCF-7 clones stably expressing control vector or a TRIM37-targeting sgRNA. β-Actin, loading control. Representative data; n = 3 biological replicates. For gel source data, see Supplementary Fig. 1. d, Representative data of a 10-day clonogenic survival of indicated MCF-7 cell lines treated with DMSO (control) or centrinone (PLK4i) (125 nM). e, Quantification of n = 3 biological replicates in d. P values, unpaired two-tailed t-test. Mean ± s.e.m. f, Immunoblot of lysates prepared from WT and TP53−/− HCT116 cells expressing a control (eGFP) or TRIM37 transgene. MCF-7 cells were used as a reference for TRIM37 protein overexpression in a 17q23-amplified cell line. β-Actin, loading control. Representative data; n = 3 biological replicates. For gel source data, see Supplementary Fig. 1. g, Representative data of a 14-day clonogenic survival assay of HCT116 cells treated with DMSO (control) or centrinone (PLK4i) (125 nM). h, Quantification of n = 3 biological replicates in g. P values, unpaired two-tailed t-test. Mean ± s.e.m. i, Immunoblot showing doxycycline-induced GST or TRIM37 expression in PLK4AS TP53−/− RPE-1 cells. β-Actin, loading control. Representative data; n = 3 biological replicates. For gel source data, see Supplementary Fig. 1. j, Representative data of a 14-day colony survival assay of PLK4AS TP53−/− RPE-1 cells expressing doxycycline-inducible GST (control) or TRIM37 transgenes, treated with DMSO (control) or 3MB-PP1 (3MB). AS, analogue sensitive. k, Quantification of n = 3 biological replicates in j. P values, unpaired two-tailed t-test. Mean ± s.e.m. l, Representative images of PLK4AS TP53−/− RPE-1 cells in j. Scale bars, 100 μm.
