Extended Data Fig. 8: Crisis cells display cytosolic DNA species.
From: Autophagic cell death restricts chromosomal instability during replicative crisis
a, Top, representative confocal microscopy images of crisis IMR90E6E7 cells (PD108) expressing RFP–NLS or immunostained with antibodies against lamin A or lamin B1. Bottom, grouped stacked bars showing the ratio of positive and negative micronuclei for each of the indicated stains. n shows number of micronuclei analysed. b, Top, representative confocal microscopy images of crisis IMR90E6E7 cells (PD108) immunostained with mitotracker dye. Bottom, grouped stacked bars showing the ratio of positive and negative cytosolic DNA products for mitotracker staining. n shows number of cytosolic DNA products analysed. c, Top, representative confocal microscopy images of crisis IMR90E6E7 cells, growing IMR90E6E7 cells expressing shRNA targeting TRF2 (day 6 post-transduction) or growing IMR90E6E7 cells expressing wild-type TRF1–FokI (48 h post-induction). The corresponding population doublings are indicated. DAPI staining in blue, telomeres in green. Bottom, grouped stacked bars showing the ratio of positive and negative micronuclei for telomeres. n shows number of micronuclei analysed. d, Top, representative confocal image of U2OS cells displaying extrachromosomal telomeric repeat (ECTR) DNA. Bottom, scatter plot with bars showing the percentage of IMR90E6E7 and U2OS cells positive for ECTRs. Growing and crisis IMR90E6E7 cells, growing IMR90E6E7 cells expressing shRNA targeting TRF2 (day 6 post-transduction) and growing IMR90E6E7 cells expressing wild-type TRF1–FokI (48 h post-induction) were used. Bars represent mean. The corresponding population doublings are indicated. Two independent experiments were performed. n shows number of cells analysed. For gel source data, see Supplementary Fig. 1.
