The effects of myeloid-specific Lipa overexpression on the transcriptomic profile of aortic plaque macrophages in mice on the Ldlr-/- background and fed a Western diet for 16 weeks

We analyzed the transcriptomic profile of aortic macrophages directly sorted from atherosclerotic aortas of two groups of mice fed a Western diet for 16 weeks: (1) mice with LysMCre-mediated myeliod-specific Lipa overexpression on the Ldlr-/- background; (2) LysMCre-negative littermates on the Ldlr-/- background. We generated a knock-in (KI) mouse model for Cre-loxP–mediated overexpression of Lipa (NM_001111100) on a C57BL/6 background by targeting the Rosa26 locus. Mice heterozygous for the insertion (LipaKI/WT) were bred to obtain homozygous LipaKI/KI mice. We bred LipaKI/KI, Ldlr-/- mice with LysMCre+/-, Ldlr-/- mice, generating mice on an atherosclerosis-prone Ldlr-/- background to assess the impact of myeloid Lipa overexpression on atherosclerosis. The LysMCre+/-, LipaKI/WT, Ldlr-/- mice (M-LipaKI) demonstrated myeloid-specific Lipa overexpression. LysMCre-/-, LipaKI/WT, Ldlr-/- (Ctrl) littermates were used as controls. Whole aortas were harvested from Ctrl and M-LipaKI mice fed a Western diet for 16 weeks and subsequently enzymatically digested into single cells. Viable aortic macrophages (DAPI-CD45+CD11b+CD64+) were sorted and subjected to low-input RNA-seq. n = 4 biological replicates per genotype, with each replicate comprising pooled samples from 2-3 male mice, resulting a total of 10 male mice per genotype.