Fig. 3. Masculinization of ATX-2-depleted germlines is caused by continuous repression of tra-2 mRNA translation. (A-C) tra-2(gf);atx-2(RNAi) gonad after staining with DAPI (A,B) and mAb1CB4 (C). Sperm are absent (C, compare with Fig. 1E) and the oocyte-like germ cells are present (boxed region in A, magnified in B). Each arrowhead in B, and in the wild-type control shown in D, indicates a single oocyte nucleus with multiple highly condensed chromosomes (irregular bar-like objects). (E,F) gld-1;atx-2(RNAi) gonad showing the presence of sperm. Boxed area in E is shown at higher magnification in F; sperm nuclei (arrowheads) are small and highly condensed. Some gld-1;atx-2(RNAi) germlines showed mitotic proliferation proximal to the sperm; this phenotype was also observed in male or masculinized hermaphrodite germlines depleted of GLD-1 at 25°C. (G-I) Nomarski micrographs of fog-2 and fog-2;atx-2(RNAi) animals with the gonads outlined in black. The gonads of most fog-2;atx-2(RNAi) animals, as shown here, were small and contained only oocytes similar to those in fog-2 gonads (arrows). Boxed area in H is shown at higher magnification in I; the spermatheca (arrowhead) does not contain any sperm. A minority of fog-2;atx-2(RNAi) animals contained gonads with either both sperm and oocytes, or only sperm. Scale bars: 50 µm.