library_construction_protocol: 60,000 L1 larvae were plated to 150 mm NGM plates supplemented with 2 mL of concentrated OP50 food. Approximately 64 hours after plating L1 larvae, day 1 adults with eggs were collected from plates using M9 solution. Harvested animals were washed twice with M9 and once with water. Animals were then suspended in TRI Reagent solution (Sigma) and frozen at -80°C until RNA extraction was performed. To extract RNAs animals suspended in TRI Reagent solution were thawed and lysed using a Bead Mill (Thermo Fisher Scientific). Bromo-chloropropane (Sigma) was added to the lysed worms and centrifuged at 12,000 x g for 15 minutes. Following centrifugation, the aqueous phase was pipetted to an equal volume of cold isopropanol and placed at -20°C for 1 hour to precipitate RNAs. RNAs were pelleted by centrifugation at 20,000 x g for 15 minutes at 4°C. Total RNAs were washed with 75% EtOH twice and suspended in RNAse free water. Small RNAs were enriched from total RNAs using the MirVana miRNA isolation kit according (Thermo Fisher Scientific). Small RNAs were treated with RNA phosphatase PIR-1 to remove gamma and beta phosphates from the 5' triphosphorylated RNAs (50). Monophosphorylated RNAs were ligated to 3' adapters ( rAppAGATCGGAAGAGCACACGTCTGAACTCCAGTCA/3ddC/, IDT) using T4 RNA ligase 2 in 25% PEG 8000 (NEB) at 15 C overnight. A 5' adapter (rArCrArCrUrCrUrUrUrCrCrCrUrArCrArCrGrArCrGrCrUrCrUrUrCrCrGrArUrCrU, IDT) was then ligated to RNAs to the product using T4 RNA ligase 1 (NEB) for 4 hours at 15 C. Ligated products were reverse transcribed using SuperScript IV Reverse Transcriptase (Thermo Fisher Scientific) to convert RNA to cDNA libraries. cDNA libraries were amplified by PCR and subsequently sequenced on an Illumina Novaseq platform (SP2 x 50 bp) at the OSU Comprehensive Cancer Center genomics core.