Histone deacetylase HDA-5 regulates lipid metabolism through H4K5 and H4K8 acetylation in Caenorhabditis elegans

To identify genes were involved in hda-5 regulation. The C. elegans N2 were subjected to hda-5 RNAi and empty vector RNAi until reach to adult. Then the worms were collected and washed by M9 buffer for three times following re-suspended in PBS, and frozen immediately in liquid nitrogen for RNA sequencing. RNA sequencing was carried out by Biomarker Technologies (BMK) GmbH (Beijing, China). Total RNA of worms were extracted by using RNeasy Plus Mini Kit (Qiagen) and assessed for quality by NanoDrop, Qubit 2.0 and Agilent 2100. mRNA was enriched via Oligo(dT) magnetic beads, fragmented, and reverse-transcribed to cDNA with random hexamers. After end repair, A-tailing, adapter ligation and PCR amplification, libraries were qualified by Qubit 2.0, Agilent 2100 and Q-PCR, then sequenced on Illumina NovaSeq 6000 (PE150). The C. elegans N2 were subjected to hda-5RNAi and empty vector RNAi until reach to adult. Total RNA from C. elegans was extracted with RNeasy Plus Mini Kit (Qiagen), assessed for quality by NanoDrop, Qubit 2.0 and Agilent 2100. mRNA was enriched via Oligo(dT) magnetic beads, fragmented, and reverse-transcribed to cDNA with random hexamers. After end repair, A-tailing, adapter ligation and PCR amplification, libraries were qualified by Qubit 2.0, Agilent 2100 and Q-PCR, then sequenced on Illumina NovaSeq 6000 (PE150). Raw reads were trimmed with trim_galore, mapped to hg38 using HISAT 2.2.1, and counted in genes by featureCounts. Normalized to CPM, differential expression was analyzed with DESeq2 (≥10 reads in at least one condition; adjusted P<0.01, |log2FC|≥1)