scRNA-seq in adipocyte progenitors in Azin-/- and wild-type mice

AZIN2 regulates adipocyte progenitor differentiation. Here we investigated how AZIN2 deficiency affects the profile of adipocyte progenitors. To this end, Azin2-/- and wild-type male mice were fed for 8 weeks a high-fat diet, subcutaneous adipose tissue was isolated and single-cell RNA-seq was performed in CD45-CD31- cells of the stromal vascular fraction. Overall design: Azin2-/- and wild-type littermate mice were fed for 8 weeks a high-fat diet, then subcutaneous adipose tissue was excised, incubated for 45 min with collagenase type I (2 mg/ml/g tissue) at 37 °C under continuous rotation. Cells were then passed through a 100-µm pore-sized cell strainer and centrifuged at 300 g for 10 min. Erythrocytes were removed from the SVF through lysis with a red blood cell lysis buffer (Biolegend) and CD31-CD45- cells were isolated using anti-CD31 and anti-CD45 MicroBeads (Miltenyi Biotec), according to manufacturer's instructions. Two mice were pooled per genotype. 10,275 wt and 11,004 Azin2-/- cells were FACS sorted.