Serum metabolites (cholesterol, triglycerides, glucose, free fatty
acids) were assayed from blood obtained from mice after 6-hour fast per
manufacturer’s protocols (Thermo Scientific TR13421, TR22421, TR15408,
and Wako Diagnostics HR Series NEFA-HR(2)).
For glucose tolerance test, mice were fasted for 6 hours, injected with
10% D-glucose (1 g/kg), and tail vein blood obtained at different time points
was analyzed using a glucometer (Contour, Bayer Healthcare, Mishawaka, IN).
Serum L-amino acid were measured from mice fed standard or high protein
Western diets for 2 month or from fasted mice given an oral protein gavage for
the indicated time points using a colorimetric L-amino acid assay kit (Abcam,
ab65347) following the manufacturer’s protocol.
Quantification of atherosclerosis at the aortic root was as
follows12 (link),21 (link). PBS-perfused hearts were placed in a
cryostat mold containing tissue freezing medium. 10 μm thick sections
were taken from the samples beginning just caudal to the aortic sinus and
extending into the proximal aorta. Slides were fixed with 4% paraformaldehyde
and stained with Oil Red O. Images were taken by EVOS XL Core Cell Imaging
system and Oil Red O positive regions were quantified using ZEN microscope
software (Carl Zeiss AG).
acids) were assayed from blood obtained from mice after 6-hour fast per
manufacturer’s protocols (Thermo Scientific TR13421, TR22421, TR15408,
and Wako Diagnostics HR Series NEFA-HR(2)).
For glucose tolerance test, mice were fasted for 6 hours, injected with
10% D-glucose (1 g/kg), and tail vein blood obtained at different time points
was analyzed using a glucometer (Contour, Bayer Healthcare, Mishawaka, IN).
Serum L-amino acid were measured from mice fed standard or high protein
Western diets for 2 month or from fasted mice given an oral protein gavage for
the indicated time points using a colorimetric L-amino acid assay kit (Abcam,
ab65347) following the manufacturer’s protocol.
Quantification of atherosclerosis at the aortic root was as
follows12 (link),21 (link). PBS-perfused hearts were placed in a
cryostat mold containing tissue freezing medium. 10 μm thick sections
were taken from the samples beginning just caudal to the aortic sinus and
extending into the proximal aorta. Slides were fixed with 4% paraformaldehyde
and stained with Oil Red O. Images were taken by EVOS XL Core Cell Imaging
system and Oil Red O positive regions were quantified using ZEN microscope
software (Carl Zeiss AG).