ZFIN ID: ZDB-FIG-130411-12
Boglev et al., 2013 - Autophagy induction is a tor- and tp53-independent cell survival response in a zebrafish model of disrupted ribosome biogenesis. PLoS Genetics   9(2):e1002379 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Day 5 to Days 7-13

Fig. 6 (A) Western blot analysis of lysates of ttis450 larvae (72 hpf) that had been injected at the 1–4 cell stage with an antisense morpholino oligonucleotide (MO) targeted to the start codon of atg5 mRNA reveals decreased levels of LC3II compared to untreated and vehicle controls, both in the presence and absence of chloroquine. Data are represented as mean +/ SD, *p<0.05. (B) Survival curve of untreated WT and ttis450 larvae compared to WT and ttis450 larvae that had been injected at the 1–4 cell stage with vehicle or atg5 MO (n>85 larvae per group). The lifespan of WT embryos/larvae is completely unaffected by injection with the atg5 MO since all three groups of WT larvae (untreated, vehicle-treated and atg5 MO-treated) progress normally through the first 10 days of development, when the experiment was terminated. The horizontal line represents untreated WT embryos (maroon squares), vehicle-injected WT embryos (green triangles) and atg5 MO-injected WT embryos (blue triangles). In contrast, ttis450 embryos respond to microinjection of the atg5 MO by impaired survival. Whereas all untreated (yellow diamonds) or vehicle-injected (purple circles) ttis450 larvae are still alive at 7 dpf, all the atg5 MO-injected ttis450 larvae are dead at this time-point (red squares). Indeed, 20% of the atg5 MO-injected ttis450 larvae have already succumbed by 3 dpf. (C–F) TEMs of WT (C) and ttis450 larvae at 120 hpf (D–F), injected at the 1–4 cell stage with the atg5-targeted MO. Inhibiting autophagy in ttis450 larvae results in the appearance of detached and shrunken IECs in the intestinal lumen (black arrow in D, E and F [boxed area in D]) but has no impact on WT IECs (C). Scale bars = 10 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
map1lc3b Ab3-map1lc3b pwp2hs450/s450 control Protruding-mouth whole organism WB
Ab3-map1lc3b pwp2hs450/s450 + MO1-atg5 standard conditions Protruding-mouth whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
pwp2hs450/s450 + MO1-atg5 standard conditions Day 5 intestinal epithelium cell detached from intestine, abnormal
Day 5 intestine cell hypotrophic, abnormal
Days 7-13 whole organism dead, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.