Sex and Molecular Differences in Cardiovascular Parameters at Peak Influenza Infection in Mice

Female and male C57BL/6J mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA) at 16 weeks age. Mice were intranasal infected with a sublethal does of 7.5×104 EID50 A/Puerto Rico/8/34 influenza virus (PR8; Charles River, Wilmington, MA, USA) or mock infected (naïve) with 1X PBS. After euthanasia at 7 days post infection lung and nasal septum were collected from animals to perform transcriptome analysis by RNA sequencing. Overall design: RNA was isolated by TRIzol Reagent (Invitrogen Life Technologies, Grand Island, NY, USA) and purified with RNeasy Mini Kit (QIAGEN; Hilden, Germany) according to manufacturer's instructions. RNAseq was used to analyze lung and nasal septum transcriptomes between females (naïve [n = 6] and infected [n = 6]) and males (naïve [n = 6] and infected [n = 6]). RNA was evaluated for quality (Qubit fluorimeter) and fidelity using Qiagen QIAxcel Advanced System, with all sample demonstrating and RIS > 8. Libraries were generated with TruSeq stranded mRNA kit (Illumina; San Diego, CA, USA), indexed with TruSeq RNA CD indexes (Illumina; San Diego, CA, USA), and sequenced with NextSeq 2000 P3 (PE100) flow cell (Illumina; San Diego, CA, USA)