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Fig. 3. FIGL-1 is required for timely mitotic progression in the germ line. (A) L3 larvae were injected with figl-1 dsRNA and the resulting embryonic lethality and sterility of their F1 progeny was quantified and expressed as a percentage (%) of the total number (n) of progeny analyzed. Although the embryonic lethality is low, the percentage of sterile progeny reaches 89%. (B) The gonadal arms at the tail end of wild-type (left image), figl-1(RNAi)-depleted progeny and figl-1(tm808)-homozygous animals (right image) were visualized within the worm by Hoechst staining. The stars mark the start of the `mitotic gonad' in the figl-1(RNAi) progeny, and the arrows indicate the mitotic and meiotic progression in the U-shaped gonad. The few nuclei that were left in the figl-1(RNAi) and figl-1(tm808) progeny all display abnormal sizes and structures (insets). Bar, 10 µm. (C) figl-1(RNAi) animals express the distal-tip cell marker Lag2::GFP. GFP expressed by a Lag-2::GFP transgene (arrows) is visualized with a fluorescent microscope in control (bottom left) or figl-1(RNAi) animals (bottom right). Upper panels show animals visualized by bright-field microscopy. Bar, 100 µm. (D,E) The number of mitotic nuclei was determined in gonads of wild-type (WT) and figl-1(RNAi) animals by immunofluorescence with antibodies against phosphorylated histone H3 (D, left images). The DNA was visualized by Hoechst staining (D, right images). The result was quantified and plotted as a percentage (%) of mitotic cells per gonad (E). Note that the number of nuclei with a positive phospho-H3 signal is higher in figl-1(RNAi) animals (P=1.17x10^–7).

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