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Fig. 8 Effect of 5-FU on the stability of TS protein in zebrafish embryos.
Healthy embryos at the 1–2 cell stage were treated with 10 μg/mL cycloheximide and 0.5 μg/mL 5-FU and incubated for another 4, 8, 16, 24 and 36 h. Embryos (n = 100) that were untreated or treated with cycloheximide and 5-FU were collected and the total protein was extracted for Western blot analysis as described in the Materials and Methods section. A, TS level in the zebrafish embryo at various time points after 5-FU treatment. TS level in the absence (circles) or presence of 5-FU (box) was analyzed by Western blot and quantified using a Hewlett Packard ScanJet 4P and NIH image 1.51 software. B, Western blot analysis showing the amount of TS at specific times after treatment with cycloheximide. Zebrafish TS embryos were left untreated (panel 1) or treated with 5-FU (panel 2) for 2 h, and 10 μg/mL cycloheximide was added to incubated for another 0 (lane 1), 4 (lane 2), 8 (lane 3), 16 (lane 4), 24 (lane 5) and 36 h (lane 6). Western blots were performed as described in the Materials and Methods section.