PGLYRP-5 and PGLYRP-2 Are Expressed in the Developing Embryos, and PGLYRP-5 Is Essential for In Vivo Protection against Infection (A–C) Cell lysates from eggs or from embryos at the indicated developmental stage were analyzed by immunoblotting with PGLYRP-2 (A) or PGLYRP-5 (B and C) antibody. Size markers are shown on the left. The results are from one of three similar experiments. (D–K) Eggs at the two- to eight-cell stage were injected with 1 nl of 0.5 mM PGLYRP-5 morpholino (P5 Mo) or a control morpholino (C Mo). At the onset of circulation (∼30 hpf), embryos were injected with S. enterica (D and E) or B. subtilis (F and G) or with S. enterica (H and I) or B. subtilis (J and K) and PGLYRP-5 protein (P5 protein) or buffer. Embryos were assayed for survival at 4 hpi or lysed for determining numbers of recovered bacteria per embryo at 2–3 hpi. The data in each panel are from two to three experiments with the following total numbers of embryos with C Mo and P5 Mo or - and + P5 protein, respectively: (D), 18 and 24; (E), 35 and 43; (F), 24 and 19; (G), 24 and 21; (H), 16 and 9; (I), 16 and 12; (J), 16 and 15; (K), 20 and 28. The survival and the numbers of recovered bacteria were significantly different at p = 0.0002 (*) or p < 0.0001 (**) between PGLYRP-5 morpholino- and control morpholino-injected groups or between PGLYRP-5 protein-injected and buffer-injected groups. The bactericidal results are means ± SE from two to three experiments.
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