Fig. S4

AID morphants, zMbd4 morphants, and Gadd45α morphants show neurogenesis defects
(A) Gross morphological defects are shown by a bright field image of AID, Gadd45α and zMbd4 morphants at 24 hpf. These defects can be reversed to a large extent by injection of mRNAs encoding the indicated proteins that are refractory to the morpholino. AID and Gadd45α mRNA were of zebrafish origin, whereas hMbd4 mRNA was of human origin. AID Scr morpholino and MBD4 Scr morpholino injected embryos showed wild type morphology (not shown). (B-C) Whole mount in situ hybridizations were performed for neurogenin-1 (B; a proneural marker) and sox-2 (C; a gene expressed in neural progenitors) on AID, Gadd45α and zMbd4 morpholino (4pg each) injected embryos or wild type embryos at 80% epiboly (a stage when wild type and morphant embryos look relatively similar in gross morphology). Note the complete absence of neurogenin-1 expression and drastic reduction in sox-2 expression in these embryos. These defects can be compensated by injection of corresponding mRNAs (refractory to the morpholino) as indicated in panel A. zAID and zGadd45a mRNA were injected at 25pgs each whereas hMBD4 mRNA was injected at 50pgs for rescue of respective morphants. AID Scr morpholino and MBD4 Scr morpholino injected embryos showed wild type expression for ngn-1 and sox-2 (not shown).