Figure 7. 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 Western blots using anti-PGLYRP-2 (A) or anti-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 2-8 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, E) or B. subtilis (F, G) or with S. enterica (H, I) or B. subtilis (J, 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/embryo at 2-3 hpi. The data in each panel are from 2-3 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 PGLYRP-5 protein-injected and buffer-injected groups.