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. 2020 Apr 27;9:e56731. doi: 10.7554/eLife.56731

Figure 3. simr-1 mutants have a transgenerational fertility defect at elevated temperature.

(A) Brood size was scored for a single generation at 20°C, followed by 11 generations at 25°C, demonstrating that simr-1 mutants become progressively sterile at 25°C. 10 broods were scored for each genotype at each generation. (B) Brood sizes for simr-1 mutant and wild-type animals were scored for 10 generations after returning to 20°C, following 10 generations at 25°C, demonstrating restoration of fertility at permissive temperature. 10 broods were scored for each genotype at each generation. (C) Wild-type and simr-1 mutant males were raised either at 20°C, a single generation at 25°C, or following 10 generations of growth at 25°C, and then mated to fog-2 females raised at 20°C. Brood sizes were scored for 10 fog-2 females, each mated to four males of the indicated genotypes, and demonstrating that simr-1 male fertility is compromised at 25°C. (D) Wild-type and simr-1 mutant hermaphrodites were raised either at 20°C, a single generation at 25°C, or following 10 generations of growth at 25°C, and then mated to four pgl-1::gfp males raised at 20°C. Brood sizes were scored for each of 10 wild-type or simr-1 mutant hermaphrodites, mated to four pgl-1::gfp males. Only plates with GFP positive progeny were scored. These data indicate that oogenesis of simr-1 is compromised after multiple generations at 25°C. (E) Number of apoptotic germ cells were counted in a minimum of 20 wild-type and simr-1 mutant gonads using CED-1::GFP engulfment as a marker for apoptotic germ cells. Animals were raised either at 20°C, or for one, two, four, seven, 10 or 11 generations at 25°C, and imaged approximately 24 hr after the L4 larval stage. Error bars indicate SEM. n.s. denotes not significant and indicates a p-value>0.05, * indicates a p-value≤0.05, *** indicates a p-value≤0.001, **** indicates a p-value≤0.0001. See Supplementary file 8 for more details regarding statistical analysis.

Figure 3—source data 1. Data used to generate Figure 3A and Figure 3—figure supplement 1.
Figure 3—source data 2. Data used to generate Figure 3B.
Figure 3—source data 3. Data used to generate Figure 3C.
Figure 3—source data 4. Data used to generate Figure 3D.
Figure 3—source data 5. Data used to generate Figure 3E.

Figure 3.

Figure 3—figure supplement 1. hpo-40 does not contribute to the progressive sterility of simr-1 mutants.

Figure 3—figure supplement 1.

Brood sizes of wild-type, hpo-40 single mutants, and simr-1; hpo-40 double mutants were scored for a single generation at 20°C, followed by 11 generations at 25°C, demonstrating that hpo-40 does not contribute to the progressive sterility of simr-1 mutants at 25°C. 10 broods were scored for each genotype at each generation.