Fig. 4

Fig. 4. Cell apoptosis was activated by kars1 loss of function through p53 pathway.

(a–c) Representative images of sibling, kars1^−/−, and kars1^−/− + p53 MO at 3 dpf after TUNEL staining. Red circle indicates eye region. Scale bars = 100 µm. (d) The eye size measurements of sibling, kars1^−/− and kars1^−/− + p53 MO at 3 dpf. (e) The expression levels of p53 pathway genes were examined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) after p53 MO injection. (f) Those downregulated genes in RNA-seq data were examined by RT-qPCR after p53 MO injection. For (e,f), the expression levels were normalized to 18 S housekeeping gene. Error bars = mean ± SD. Two-tailed unpaired Student’s t-test with Holm–Šídák multiple comparisons correction: ns, not significant p ≥ 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001. (g) Representative images of uninjected control, kars1 F₀ mutant, kars1;tp53 F₀ mutant, and kars1 F₀ mutant coinjected with kars1 messenger RNA (mRNA) at 5 dpf. Lateral view, anterior to the left. (h,i) Measurement of eye and head size in control, tp53 F₀ mutant, kars1 F₀ mutant, kars1;tp53 F₀ mutant, and kars1 F₀ mutant coinjected with 150 picogram (pg) and 200 pg of kars1 mRNA at 5 dpf. For (d,h,i), each symbol represents one animal. Error bars = mean ± SD. One-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test: ns, not significant p ≥ 0.05 and ****p < 0.0001. (j) Kaplan–Meier survival curves. Time is shown in days. Log rank test: ****p < 0.0001. (k) Histology analysis of uninjected control, kars1 F₀ mutant and kars1;tp53 F₀ mutant by cross-section.