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Fig. 3 Rescue of neural apoptosis and MHB marker loss in grhl2b morphants by eng2a mRNA. (A-F) Rescue of cell death (arrows) shown by bright-field microscopy (A-C) and TUNEL assay (D-F) in lateral views of 26 hpf embryos sequentially injected with MO:grhl2b and either lacZ mRNA (A,D) or eng2a mRNA (B,E). MO:control embryos injected with lacZ mRNA are shown (C,F). (G-I) Rescue of expression (arrows) of the MHB maintenance marker pax2a in 26 hpf embryos sequentially injected with MO:grhl2b and either lacZ (G) or eng2a (H). MO:control embryo injected with lacZ mRNA is shown (I). (J,K) MHB folding in grhlb2 morphants (J) is not rescued by injection of eng2a mRNA (K). Injection of lacZ (J) as control. (L,M) Cooperativity between grhl2b and eng2a ATG-blocking MOs in inducing loss of pax2a expression and apoptosis (L, arrowhead; dorsal view, inset), but not MHB folding (brackets, M). Lateral views. (N) Q-RT-PCR analysis of MHBs extracted from n=3 MO:grhl2b 24 hpf morphants and MO:control embryos showing expression of p75NTR.