Flies of the specified genotypes(w;;Mettl3[null]/+, w;;Mettl3[null]/Mettl3[cat], w;;Df(3 R)BSC461/ + , w;;Ythdf[NP3]/Df(3 R)BSC461, were aged to 1 or 3 weeks at 25 °C. Female heads were dissected and collected for total RNA extraction using TRIzol reagent. Sequencing libraries were prepared using the TruSeq Stranded Total RNA Sequencing Kit (Illumina) following the manufacturer’s protocol. Sequencing was performed on a HiSeq 2500 System in paired end read mode, with 100 bases per read at the Integrated Genomics Operation (IGO) at Memorial Sloan Kettering Cancer Center.
RNA sequencing libraries were mapped to the Drosophila reference genome sequence (BDGP Release 6/dm6) using HISAT2105 (link) under the default settings. Gene counts were obtained by assigning and counting reads to the Ensembl transcript models for BDGP6.94 using Rsubread106 (link). Differential gene expression analysis was performed with comparisons as listed in Supplementary Dataset5 using the R package DESeq2107 (link) and applying a strict adjusted p-value cutoff of 0.05.
RNA sequencing libraries were mapped to the Drosophila reference genome sequence (BDGP Release 6/dm6) using HISAT2105 (link) under the default settings. Gene counts were obtained by assigning and counting reads to the Ensembl transcript models for BDGP6.94 using Rsubread106 (link). Differential gene expression analysis was performed with comparisons as listed in Supplementary Dataset
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