Adipocyte Differentiation in BmN4 Cells

For the induction of adipocyte differentiation, BmN4 cells were cultured in IPL-41 medium supplemented with 10% fetal bovine serum, 10 μg/ml insulin (SIGMA), 0.5 μM 3-isobutyl-1-methylxanthine (SIGMA) and 250 μM dexamethasone (SIGMA). Medium was changed twice a week and collected 10 and 20 days after the drug addition to prepare RNA sequence libraries. These libraries were mapped to the silkworm genome and genemodel, transposon, and the novel inserts identified in this paper using Hisat2 with the default parameters, and the mapped reads were calculated by coverageBed of bedtools [41 (link), 49 (link)]. Silkworm homologs of piRNA-related genes were identified using blast of silkbase. Since the genemodel for BmAgo3 was splited into three (KWMTBOMO01223-5) and the genemodel for Tudor was splited into two (KWMTBOMO06482-3), the RPKM was calculated by combining them.