Intermittent childhood hyperlipidaemia alters resident arterial macrophages and accelerates atherosclerosis
Intermittent childhood hyperlipidaemia alters resident arterial macrophages and accelerates atherosclerosis
Aortas were harvested and cleaned of contaminated tissues. Aortas were subjected to enzymatic digestion method and single cell suspensions of aortic cells were prepared. Cell suspensions were incubated with Zombie Aqua™ Fixable Viability Kit (BioLegend) and then cells were stained with fluorescently labelled anti-mouse antibodies against CD45, CD11b and CD64. Aortic macrophages were identified as CD45+CD11b+CD64+. Animals were perfused with cold PBS to remove any trace of blood.  Aortas were harvested and cleaned of contaminated tissues under a dissection microscope 16 wks mouse high fat diet. Align to GRCm39 reference genome using star (version 2.6.1d) RNA extraction and residual DNA removal upon DNase digestion was performed with the RNeasy mini kit (Qiagen) according to the manufacturer's instructions. After quality check with Agilent Bioanalyser 2100 system, total RNA was made into sequencing libraries. Takara's SMART-Seq v4 Ultra Low Input RNA Kit.