Transcriptome analysis of mouse macrophages during Brucella abortus infection clarifies the survival mechanisms of the bacteria
Transcriptome analysis of mouse macrophages during Brucella abortus infection clarifies the survival mechanisms of the bacteria
Mouse macrophages J774A.1 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Euroclone, Italy), supplemented with 10% (v/v) fetal bovine serum (FBS, Euroclone, South America, origin EU approved), penicillin-streptomycin solution (100 U/mL penicillin and 100 µg/mL streptomycin) (Euroclone, Italy) and grown at 37°C with 5% CO2 in a humidified incubator. Total RNA from J774.1 infected with B. abortus was isolated using Trizol following the manufacturers protocol. The concentration and quality of all isolated RNA samples were measured using the TapeStation 4200 (Agilent Technologies, USA) and Qubit 2.0 fluorometer (Thermo Fisher Scientific, USA). RNA samples that exhibited RNA integrity numbers ranging from 6.6 to 10 were selected for the library preparations. Libraries were prepared using the Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus kit (Illumina, San Diego, USA) according to the manufacturers instructions, starting with 1 µg total RNA.