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Fig. S2.
Supplementary material. Expression of heterochronic genes during metamorphosis. (A-F) RT-qPCR from brain tissue during metamorphosis for (A) miR-125 (lin-4 orthologue), (B) let-7a, (C) lin28b, (D) trim71 (lin41 orthologue) and the TH target genes (E) thrβA and (F) klf9. RNA levels were normalized to 5S (A, B) and eef1α−1 (C-F) and compared to levels at st.50 (average ±SEM, n=2–5 independent experiments). *p<0.05, **p<0.01, ***p<0.001, t-test against hypothetical value =1 (no change compared to st.50). (G) Western blot analysis for Lin28a in spinal cord samples. Tubulin was used as loading control. (H, I) Western blot for Lin28b of (H) spinal cord and (I) brain samples at different stages. Upper arrow shows the full length Lin28b and lower arrow shows a putative splicing isoform. Tubulin was used as loading control. (J-M) In situ hybridization for lin28b in spinal cord and brain sections of st.50 animals using sense (s-) or antisense (as-) probes. Bar, 50 µm. (N,O) RT-qPCR from intestine during metamorphosis for (N) lin28b and (O) thrβA. RNA levels were normalized to eef1α−1 and compared to levels at st.50 (average ±SEM, n=2–3 independent experiments).
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