Fig. 2: Transcription stress in Tcea1–/– MEFs leads to genomic instability.
From: Transcription stress at telomeres leads to cytosolic DNA release and paracrine senescence
A Immunostaining of 8-Oxoguanine (8-oxoG) in untreated wt and Tcea1–/– MEFs (n = 3). The graph depicts the 8-oxoG MFI per cell nucleus for untreated, H2O2-treated and NAC-treated cells. B Serine 2 (pS2)-phosphorylated RNAPII (pS2-PolII) in whole-cell extracts from wt and Tcea1–/– MEFs. The graph depicts the total RNAPII-normalized protein expression levels (n.e.l., n = 3). C Dot blot against 8-oxoG in pS2-PolII ChIP samples from wt and Tcea1–/– MEFs. The graph represents pS2-PolII ChIP samples normalized over input samples (n.l./Input, n = 3). D BrU incorporation in untreated wt and Tcea1–/– MEFs. The graph shows the BrU MFI per cell nucleus for untreated and DRB-treated cells (n = 3). E Dot blot against s9.6 in untreated and Rnase H-treated genomic DNA from wt and Tcea1–/– MEFs. The graph represents input samples normalized over Ethidium Bromide (EtBr) labeling, as a loading control (n = 3). F Immunofluorescence detection of γH2AX and 53BP1 co-localized foci in Tcea1–/– MEFs, in the presence or absence of transfected recombinant RNase H. The graph represents the percentage of cells with >3 co-localized γH2AX/53BP1 foci per cell nucleus (n = 4). G BrdU immunostaining under non-denaturing (Non-Den) conditions in wt and Tcea1–/– MEFs. The graph depicts the BrdU MFI per cell nucleus (n = 3). Data analysis was performed using two-tailed Student’s t test. All data are presented as mean values ± SEM. Unless otherwise indicated, n = biologically independent experiments and scale bars are set at 5 μm. Source data are provided as a Source Data file.
