High-Fat Diet Feeding
Objective: To examine SIRT3 gene expression changes in mice fed a high-fat diet for three months
Protocol Steps
Animal housing and diet assignment
8-week-old male C57BL/6 mice were singly caged and assigned to either control diet or 35% fat-enriched chow groups
Note: Mice were individually housed in cages
View evidence from paper
“8-week-old male mice were singly caged. At 8 weeks of age, control mice were fed ad libitum with NIH-31 standard diet”
Diet feeding period
Mice were fed their assigned diet (control or 35% fat-enriched chow) for three months
Note: Control group received standard diet ad libitum; experimental group received 35% fat-enriched chow
View evidence from paper
“For the high-fat diet feeding experiment, 8-week-old male mice were fed a control diet or a 35% fat-enriched chow (BioServ) for three months”
Tissue collection
At the end of the three-month feeding period, various tissues were harvested from mice in both groups
Note: Tissues were collected for SIRT3 gene expression analysis
View evidence from paper
“Various tissues were also harvested from mice fed the control diet to examine SIRT3 gene expression by Western blot analysis at the termination of the study”
Western blot analysis
SIRT3 gene expression was analyzed in harvested tissues using Western blot analysis
Note: Anti-mouse SIRT3 serum raised against the C-terminus was used for high-fat diet analyses
View evidence from paper
“Various tissues were also harvested from mice fed the control diet to examine SIRT3 gene expression by Western blot analysis”