Figure EV3. gtsf‐1 animals recapitulate the phenotypes of alg‐3/4 and ergo‐1 mutants (related to Fig 3).

• A, B
  Representative pictures of progenies from synchronized parents of wild‐type (A) and gtsf‐1 mutants (B) grown at 25°C. In (A), L1 larvae and gastrulated embryos can be observed, while in (B) only unfertilized oocytes are identified.
• C
  Percentage of dead eggs and unfertilized oocytes laid by wild‐type and gtsf‐1 mutant worms when grown at 25°C. These counts were obtained from the same experiment as in Fig 3A. n = 10.
• D
  % of males is plotted for the indicated strains. gtsf‐1 mutants show a percentage of males comparable to alg‐3/4 and rrf‐3 mutants, but lower than mut‐7 mutants. Spontaneous male incidence was measured in the progenies of 20 worms. For each plate, percentage of males was calculated and the percentages of the 20 plates were subsequently averaged to obtain the final male percentage value. Error bars represent SEM.
• E
  % Survival of animals of the indicated genotypes on empty vector (EV) and pos‐1 RNAi. Per strain, progenies of n = 9–12 worm were scored for embryonic lethality. gtsf‐1 animals respond to pos‐1 RNAi identically to wild type. Error bars represent SD.
• F
  This panel shows a representative fraction of animals from the dpy‐13 RNAi experiment (shown in Fig 3C). Genotypes and treatment with either empty vector (EV) or dpy‐13 RNAi are indicated in the figure. Scale bars represent 0.5 mm.
• G
  Overview of the 22G transgenes. The control transgene consists of GFP controlled by the ubl‐1 promoter and 3′UTR sequences, which allow ubiquitous expression. The 22G‐siR‐1 sensor is similar to the control transgene except for a binding site for 22G‐siR‐1 on the 3′UTR that reports on the activity of ERGO‐1‐dependent 22G‐RNAs.
• H
  Genetic requirements for 22G‐sensor silencing. Representative fluorescence images of animals carrying 22G‐siR‐1 sensor combined with other mutations. Scale bars represent 0.5 mm.