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Figure S1. Analysis of dcap-2(tm2470) and patr-1(tm2402) deletion mutants. (A) Schematic of the WT and tm2470 dcap-2 alleles. Two DCAP-2 isoforms (a and b) consist of 806 and 786 amino acids, respectively. Only the a isoform is shown. The tm2470 deletion, which has been sequenced from an RT-PCR product, produces a truncated version of each isoform. For example, the mutant version of the a isoform would consist of the N-terminal 192 amino acids along with an additional novel 81 residues. These predicted mutant proteins would be nonfunctional because they lack most of the conserved Dcp2 domain (also known as box A; Wang, Z., X. Jiao, A. Carr-Schmid, and M. Kiledjian. 2002. Proc. Natl. Acad. Sci. USA. 99:12663–12668) and the entire catalytic MutT/NUDIX domain (Bessman, M.J., D.N. Frick, and S.F. O’Handley. 1996. J. Biol. Chem. 271:25059–25062; Wang et al., 2002). (B) The patr-1(tm2402) deletion, which was sequenced from an RT-PCR product, encodes only the N-terminal 186 amino acids (of 833 total), along with 46 unrelated residues. No PATR-1 protein expression has been detected in patr-1(tm2402) by either Western blotting (Fig. 3 C) or antibody staining (E). (C) Gonad abnormalities in homozygous dcap-2(tm2470) and patr-1(tm2402) adult hermaphrodites. Nomarski images of living animals are shown. Compared with WT (N2), the gonads of dcap-2(tm2470) and patr-1(tm2402) hermaphrodites were typically characterized by an extended pachytene region and contained fewer cellularized oocytes. dcap-2(tm2470) homozygotes were viable and generally normal in appearance but were sedentary and slow moving. patr-1(tm2402) animals depended on maternally contributed patr-1 for viability (Table S1). This lethality was prevented when patr-1(tm2402) homozygotes were mated with fog-2 males (Table S1), which indicates that either maternally or zygotically supplied patr-1 is sufficient to allow development to adulthood. Both dcap-2(tm2470) and patr-1(tm2402) mutants ceased to lay eggs during the second day of adulthood, retained many embryos, and produced fewer offspring compared with WT (P < 0.05; Table S1). (D) PATR-1 is expressed predominantly in somatic tissues. PATR-1 levels were analyzed by a Western blot of 50 WT or patr-1(tm2402) intact adult hermaphrodites and 100 WT dissected gonads (GN). (E) The absence of PATR-1(N) staining in homozygous patr-1(tm2402) embryos. These embryos lack both maternally provided and zygotic patr-1. An ∼28-cell embryo was stained with anti–CGH-1 and anti–PATR-1(N) antibodies. CGH-1 is readily detectable in the germ (P) granules and in additional somatic foci, but PATR-1(N) staining was not detectable even with a 10-fold increase in exposure (not depicted). A representative patr-1(tm2402) embryo is shown here, and WT embryos that were similarly stained are shown in Fig. 3 B. (F) Somatic P-bodies in dcap-2(tm2470) adult animals. CGH-1 (green) and PATR-1 (red) particles colocalize in many somatic cells in dcap-2(tm2470) adults. Neither CGH-1 nor PATR-1(N) staining is detectable postembryonically in the WT (Fig. 3 A; not depicted; n > 100). Dashed lines outline the head (including the pharynx) and tail (including the intestine and rectum). Single-plane confocal images are shown. Bars: (C and F) 20 µm; (E) 10 µm.