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Fig. 5. Effect of JNK inhibitor on tail regeneration. (A–H) Tail-amputated tadpole was incubated in water containing 8 μM JNK inhibitor SP600125 (B, D, F and H) or in control water containing DMSO (A, C, E and G). (E and F) Sections stained with hematoxylin and eosin. (G and H) Whole-mount in situ hybridization for Xes1. Arrowheads in (A–D) show amputation planes. Arrow in (E) indicates notochord precursor cells. White arrowhead in (G) indicates positive signal for Xes1 expression. All specimens are lateral view, anterior to the left. sc, spinal cord; nc, notochord. Scale bars in (A–D, G and H) 500 μm; (E and F) 50 μm. (I) The mean length of the regenerated tails was determined on day 5 and is indicated with the standard deviation. *P value determined with Student’s t test is less than 0.000001. (J) RNA was isolated from the amputated tail on day 0, and from the tail amputated and treated with SP600125 (8 μM) or DMSO for two days, and used for RT-PCR analysis. |
