To validate key RNA-seq findings, we re-isolated RNA from lung macrophages of infected mice and performed qRT-PCR analysis of selected genes of interest shown by RNA-seq as significantly altered between the sexes during infection, including Ighg1, Il1b, Ctsc, Cacna1c, Lamp2, Ulk1, Tubb1, Adra1a, and Col9a1 (Fig. 4c and d). This evidence concerns the gene TUBB1 and infection.