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. 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)