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Fig. S3
zTERT mRNA and telomerase activity levels during embryogenesis and in adult fish.
(A) Whole-mount mRNA in situ hybridizations were performed at the indicated stages. All embryos are lateral views with the animal pole orientated upwards (0.2-6 hpf) and the head positioned to the left (12-48 hpf). Note that the expression of TERT mRNA is absent at 6 hpf, detectable at 12 hpf, and is at high levels, particularly in the eye, midbrain, and hindbrain, by 16 to 48 hpf. Sense probes were used as a negative control.
(B) The expression of zTERT mRNA was analyzed by RT-PCR during embryogenesis and was found to be decreased by 6 hpf, at high levels during 9-24 hpf, and decreased again by 72 hpf. Similar results were obtained for a number of individual embryos (more than 10 embryos at each time point).
(C) Telomerase activity in whole zebrafish embryos during early development assayed by TRAP. Similar results were obtained for at least 5 embryos at each time point.
(D) Detection of zTERT mRNA expression by RT-PCR. Representative results from three independent experiments are shown. Similar results were obtained in three-independent experiments by using different individual samples.
(E) Detection of telomerase activity by TRAP assay in adult zebrafish (12 months old) tissues. The average values of two-independent duplicated experiments are shown in the graph. C; negative control.