Morpholinos and RNA injections
The effect of zebrafish patched1 and patched2 morphants has been previous reported (Koudijs et al., 2008) and, therefore, we ordered Morpholino Oligonucleotides (MO) targeting each translation start site of zebrafish patched1 and patched2 mRNA, as well as control MOs, were developed by Gene Tools (Gene Tools LLC) and designed as follows: the patched1 translation-blocking MO (ATG MO), 5’-AGGAGACATTAACAGCCGAGGCCAT-3’ and the patched2 ATG MO, 5’-CATAGTCCAAACGGGAGGCAGAAGA-3’. MOs were diluted in RNAse-free water to 0.1-0.5 ng/nl (ptch 1, ptch 2), 5 ng/nl (con MO).