RNA-seq of A. fumigatus WT and dRmiA mutant exposed to miltefosine
A. fumigatus WT and deltarmiA were inoculated in 50 ml of Vogels medium and grown at 37 C for 16h under agitation. The suspensions were centrifuged and washed with PBS. Mycelia was suspended in VMM media containing glucose supplemented with 3 mg/mL of miltefosine, or in the absence of any drug, and incubated at 37 C for additional 30 min. Total RNA was extracted by Trizol method. Subsequently, 10 g of total RNAs were subjected to RNA purification using DNase I (New England Biolabs Inc.) and the quality was checked on 2% agarose gel and verified using the Agilent Bioanalyser 2100 (Agilent technologies).