Journal: eLife

Article Title: Opposing p53 and mTOR/AKT promote an in vivo switch from apoptosis to senescence upon telomere shortening in zebrafish

doi: 10.7554/eLife.54935

Figure Lengend Snippet: ( A-B ) Western blot analysis of DNA damage and senescence-associated proteins in gut and testis of 3 month ( A ) or 9-month-old ( B ) of WT and tert-/- siblings (N >= 5 fish). Representative western blot (left panel) and corresponding quantification (right panel) showing induction of DNA Damage Response (H2A.X-P and p53) in 3-month-old and senescence (p15/16) in 9-month-old tert-/- zebrafish. ( C ) RT-qPCR analysis of senescence associated genes p15/16 and p21. RT-qPCR graphs are representing mean ± SEM mRNA fold increase after normalisation by rpl13a gene expression levels (* p-value<0.05; ** p-value<0.01, using the Mann-Whitney test). Figure 2—source data 1. Western Blot quantifications, as plotted in . Figure 2—source data 2. Real-time qPCR data of p15/16 and p21, as plotted in .

Article Snippet: Antibody , anti-zebrafish p53 (rabbit polyclonal) , Anaspec , #55342; RRID: AB_2287635 , WB (1:1000).

Techniques: Western Blot, Quantitative RT-PCR, Gene Expression, MANN-WHITNEY

Journal: eLife

Article Title: Opposing p53 and mTOR/AKT promote an in vivo switch from apoptosis to senescence upon telomere shortening in zebrafish

doi: 10.7554/eLife.54935

Figure Lengend Snippet: ( A and E ) Representative haematoxylin and eosin-stained sections of gut ( A ) (scale bar: 40 µm) and testis ( E ) (scale bar: 25 µm) from 6-month-old WT, tert-/-, tp53-/ - and tert-/- tp53-/ - siblings (N = 3 fish each);. Mutation of tp53 in tert-/- fish rescues short-telomere induced tissue defects. ( B and F ) Representative western blot analysis of AKT-p and SOD2 in gut ( B ) and testis ( F ) (N = 2 fish each). Mutation of tp53 in tert-/- fish prevents phosphorylation of AKT and downstream downregulation of SOD2 leading to a rescue of increased ROS levels (C and G; N = 3 fish per genotype). ( D and H ) Representative images of SA-β-GAL staining of gut (scale bar: 40 µm) ( D ) and testis (scale bar: 25 µm) ( H ) from 6 month-old WT, tert-/-, p53-/ - and tert-/- p53-/ - siblings (N = 3 fish). Data are represented as mean ± SEM (** p-value<0.01, using t-test). Figure 5—source data 1. ROS levels measurements, as plotted in .

Article Snippet: Antibody , anti-zebrafish p53 (rabbit polyclonal) , Anaspec , #55342; RRID: AB_2287635 , WB (1:1000).

Techniques: Staining, Mutagenesis, Western Blot, Phospho-proteomics

Journal: eLife

Article Title: Opposing p53 and mTOR/AKT promote an in vivo switch from apoptosis to senescence upon telomere shortening in zebrafish

doi: 10.7554/eLife.54935

Figure Lengend Snippet: Early telomere shortening triggers p53-dependent apoptosis and inhibition of cell proliferation. At early age, apoptosis is the predominant cell fate and it mostly counterbalanced by compensatory proliferation of neighboring cells. However, inhibition of cell proliferation results in a progressive loss of tissue cellularity, eventually leading to tissue damage. As age progresses, loss of tissue homeostasis triggers the pro-proliferative mTOR/AKT pathway. Akt phosphorylates FoxO, inducing its translocation from the nucleus to the cytoplasm. Loss of FoxO transcriptional activity reduces mitochondrial SOD2 expression generating mitochondria oxidative stress through increased ROS levels. Mitochondrial dysfunction eventually triggers p15/16 expression and senescence becomes the predominant cell fate.

Article Snippet: Antibody , anti-zebrafish p53 (rabbit polyclonal) , Anaspec , #55342; RRID: AB_2287635 , WB (1:1000).

Techniques: Inhibition, Translocation Assay, Activity Assay, Expressing

Journal: eLife

Article Title: Opposing p53 and mTOR/AKT promote an in vivo switch from apoptosis to senescence upon telomere shortening in zebrafish

doi: 10.7554/eLife.54935

Figure Lengend Snippet:

Article Snippet: Antibody , anti-zebrafish p53 (rabbit polyclonal) , Anaspec , #55342; RRID: AB_2287635 , WB (1:1000).

Techniques: Sequencing, Control, In Situ, Cell Viability Assay, Staining