IMAGE

Fig. 8

ID
ZDB-IMAGE-130411-17
Source
Figures for Boglev et al., 2013
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Figure Caption

Fig. 8

(A) Western blot analysis of Tp53 protein in whole cell lysates of WT (lane 1) and ttis450 (lane 2) larvae at 96 hpf reveals up-regulation of Tp53 expression in ttis450. Larvae treated with roscovotine (ROS; lane 3) to induce Tp53 protein expression or untreated larvae (lane 4) are positive and negative controls, respectively. The Actin signal provides a loading control. (B–E) Relative expression of ΔN113p53 (B), mdm2 (C), cyclinG1 (D) and p21 (E) mRNAs in WT, ttis450 (pwp2h-/-), tp53M214K/M214K (tp53-/-) and ttis450;tp53M214K/M214K (pwp2h-/-;tp53-/-) larvae at 96 hpf (n = 3) demonstrates that the expression of Tp53 target genes is increased in ttis450 compared to WT larvae (compare first 2 bars in all graphs). The Tp53 response is diminished on the tp53M214K/M214K background, as expected (compare 2nd and 4th bars). Data were normalised by reference to Elongation factor alpha (Elf-α) expression. (F) Western blot analysis of LC3 in whole cell lysates of tp53-mutant (tp53M214K/M214K) and ttis450;tp53M214K/M214K larvae at 96 hpf. The elevated autophagic flux in ttis450 larvae due to ribosomal stress is not diminished on a tp53-mutant background. (G) Graphical representation of the data shown in F and two additional experiments. Bars represent the mean +/- SD (n = 3), *p<0.05. (H) Transmission electron micrographs of IECs of ttis450;tp53M214K/M214K larvae at 120 hpf (right panel) reveal electron dense vesicles, resembling autolysosomes (white arrowhead), in comparable numbers to those found in ttis450 larvae with WT Tp53 expression (left panel).

Acknowledgments
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