Fig. S1

Fig. S1 Hh signalling does not drive cell cycle exit of adaxial cells.

Embryos from a smo^b641/+ incross, treated with cyA or injected with cdkn1c MO were exposed to BrdU (or not) at ~ 6 ss at 4 °C for 20 min, transferred to Embryo Medium at 28.5 °C for 30 min, and BrdU was detected by immunohistochemistry at ~ 8 som. (A). Smo^b641 mutants were genotyped by in situ hybridization for myod and separated prior to BrdU staining. As a control, omission of BrdU prevented all nuclear labelling. Note the lack of BrdU in smo^b641 mutant adaxial cells lacking myod mRNA, but its presence in more anterior notochord, consistent with Fucci results showing cells in S/G2/M in maturing notochord (Sugiyama, M., Sakaue-Sawano, A., Iimura, T., Fukami, K., Kitaguchi, T., Kawakami, K., Okamoto, H., Higashijima, S.I., and Miyawaki, A. 2009. Illuminating cell-cycle progression in the developing zebrafish embryo. Proc Natl Acad Sci U S A 106: 20812-20817.) (B). hsp90a mRNA reveals adaxial cell nuclei. Note the lack of BrdU⁺ nuclei in the hsp90a-containing adaxial cells and in the notochord/myod focal plane in the high magnifications at right, irrespective of genotype/treatment. Bars: 25 μm.