Hif1a-WT
Mus musculus
Protocols: Aortic arches were harvested from male mice with smooth muscle cell–specific deletion of Hif1a (Myh11-CreERT2; Rosa26-tdTomato; Hif1a-floxed; Hif1aSMC-KO) and Cre-positive wild-type controls. Atherosclerosis was induced by tail-vein injection of rAAV8-PCSK9 at 9 weeks of age followed by 21 weeks of high-fat diet. Arches were pooled by genotype (Hif1aWT n=6, Hif1aSMC-KO n=8), cleaned in situ of fat and surrounding tissue, and pretreated with 2 mg/ml collagenase II in HBSS for 15 min at 37°C to remove the adventitia. Stripped arches were washed, opened longitudinally, cut into small pieces, and digested in HBSS containing 200 µg/ml hyaluronidase, 250 µg/ml Liberase, and 200 µg/ml DNase for 60 min at 37°C. The suspension was filtered through a 70 µm strainer, stained with DAPI (1 µg/ml) and Draq5 (5 µM), and viable cells (Draq5+/DAPI–) were isolated on a FACSAria cell sorter. Viability (>80%) was confirmed on a Countess III counter. Mice were group-housed under specific pathogen-free conditions with a 12-h light–dark cycle at 23 ± 1°C and 50 ± 5% humidity, with ad libitum access to water and food (SAFE D184 when not on high-fat diet) at CNIC. Only male offspring were studied, as the Myh11-CreERT2 transgene is inserted on the Y chromosome. Cre recombination was induced by intraperitoneal injection of tamoxifen (2 mg in corn oil) on five consecutive days from 6 weeks of age. To induce atherosclerosis, mice received a tail-vein injection of rAAV8-PCSK9 (1×10¹¹ vector genomes) at 9 weeks of age followed by high-fat diet (SSNIFF S9167-E011) for 21 weeks. Aortic arches were harvested at 30 weeks of age for single-cell analysis. Up to approximately 10,000 sorted viable cells per sample were loaded onto a Chromium Next GEM Chip G and partitioned into Gel Beads-in-Emulsion using the 10x Genomics Chromium controller. Cells were lysed within GEMs and mRNA was captured and reverse-transcribed into barcoded cDNA using the Chromium Next GEM Single Cell 3′ Kit v3.1 (10x Genomics) according to the manufacturer's instructions. Following cDNA amplification, 3′ gene expression libraries were constructed using the Chromium Next GEM Single Cell 3′ Kit v3.1 (10x Genomics) according to the manufacturer's instructions. Library size distribution was assessed on a 2100 Bioanalyzer (Agilent) and library concentration was quantified using a Qubit fluorometer (Thermo Fisher Scientific).
ENA first public
2026-06-21
INSDC center name
Atherosclerosis Research Unit, Department of Clinical Medicine, Aarhus University, Aarhus Denmark
INSDC status
public
Submitter Id
E-MTAB-17142:Hif1a-WT
age
30
broker name
ArrayExpress
cell type
vascular smooth muscle cell
collection date
not collected
common name
house mouse
developmental stage
adult
diet
High-fat diet (SSNIFF S9167-E011) for 21 weeks to induce atherosclerosis, following rAAV8-PCSK9 injection at 9 weeks of age. Standard chow (SAFE D184) prior to high-fat diet.
disease
atherosclerosis
genotype
Myh11-CreERT2; Rosa26-tdTomato; Hif1a^wt/wt
geographic location (country and/or sea)
not collected
individual
pool
isolate
not applicable
organism part
thoracic aorta
sample name
E-MTAB-17142:Hif1a-WT
scientific_name
Mus musculus
sex
male
strain
C57BL/6J