Fig. 5

Anxa4 knockdown causes stage and tissue specific apoptosis in the developing foregut. (a)-(c) Bright field microscopy of 31 hpf standard control MO (a), Anxa4 MO1 (b) and Anxa4 MO2 (c) morphant zebrafish embryos showing a mild developmental delay in the Anxa4 morphants. (d)-(f′′) The ventral views of 31 hpf Tg(sox17:GFP) foregut stained for Anxa4 and cleaved Caspase 3 (*Casp 3). In Anxa4 morphants (MO1 in (e)-(e′′); MO2 in (f) and (f′′), but not in control morphants (d) and (d′′), Anxa4 labeling is mostly lost (e) and (f) and patches of GFP⁺ apoptotic blebs can be found restricted to the Anxa4 morphant liver buds (white arrowheads). A subset of these GFP blebs are also positive for cleaved Caspase 3 (yellow arrowheads). (g) Graph summarizing relative incidence of liver apoptosis in anxa4 morphants, which peaks at 31 hpf. (h) Graph of a representative set of real-time RT-PCR results showing that in 27 hpf Anxa4 MO1 morphants, there is elevated expression of caspase 8 and Δ113p53.