Figure 5. GTSF‐1 interacts with RRF‐3 in the adult germline, independently of RNA .

• A, B
  Volcano plots representing label‐free proteomic quantification of GTSF‐1 IPs from adult worm extracts. For each strain, IPs were performed and measured in quadruplicates. Log₂ fold enrichment of individual proteins in one strain versus another is given on the x‐axis. The y‐axis indicates the Log₁₀‐transformed probability of the observed enrichments (see Materials and Methods for details). Proteins in the background are represented as green dots, while orange dots show enriched proteins. In (A), GTSF‐1 was immunoprecipitated using our polyclonal anti‐GTSF‐1 antibody (in wild‐type and gtsf‐1 mutant worms), while in (B), an anti‐FLAG antibody was used to pull‐down GTSF‐1::mCherry::3xFLAG (in wild‐type and strains carrying the rescuing transgene).
• C
  To test interaction between GTSF‐1 and RRF‐3 in adult worms by Western blot, GTSF‐1::HA was pulled‐down via HA immunoprecipitation. Interaction was also tested in the presence/absence of ERI‐5 by introducing an eri‐5(tm2528) mutation in the background. Multi‐channel secondary antibody detection was performed with an Odyssey CLx apparatus (see Materials and Methods). For the anti‐GTSF‐1, 1 represents GTSF‐1::HA and 2 represents untagged GTSF‐1.
• D
  Testing RNA dependency on the interaction between GTSF‐1 and RRF‐3 by adding RNase. Extracts from adult worms were used. Secondary antibody detection was performed with the Odyssey CLx setup.